Thrombin inhibitors

ABSTRACT

Novel five-membered heterocyclic amidines, their preparation and use as competitive inhibitors of trypsin-like serine proteases, especially thrombin and kininogenases such as kallikrein. Pharmaceutical compositions which contain the compounds as active ingredients, and use of the compounds as thrombin inhibitors, anticoagulants and antiinflammatory agents.

The present invention relates to novel five-membered heterocyclicamidines, to their preparation and to their use as competitiveinhibitors of trypsin-like serine proteases, especially thrombin andkininogenases such as kallikrein. The invention also relates topharmaceutical compositions which contain the compounds as activeingredients, and to the use of the compounds as thrombin inhibitors,anticoagulants and antiinflammatory agents.

Thrombin belongs to the group of serine proteases and plays a centralpart in the blood coagulation cascade as terminal enzyme. Both theintrinsic and the extrinsic coagulation cascade lead via a plurality ofamplifying stages to the production of thrombin from prothrombin.Thrombin-catalyzed cleavage of fibrinogen to fibrin then initiates bloodcoagulation and aggregation of platelets which, in turn, due to thebinding of platelet factor 3 and coagulation factor XIII, and a largenumber of highly active mediators, enhance thrombin formation.

The formation and action of thrombin are central events in thedevelopment both of white, arterial and of red, venous thrombi and aretherefore potentially effective points of attack for drugs. Thrombininhibitors are, by contrast with heparin, able independently ofcofactors completely to inhibit simultaneously the effects of freethrombin and of that bound to platelets. They are able to prevent in theacute phase thromboembolic events after percutaneous transluminalcoronary angioplasty (PTCA) and lysis, and to act as anticoagulants inextracorporeal circulation (heart-lung machine, hemodialysis). They canalso be used generally for the prophylaxis of thrombosis, for exampleafter surgical operations.

It is known that synthetic arginine derivatives influence the enzymaticactivity of thrombin by interacting with the active serine residue ofthe protease thrombin. Peptides based on Phe-Pro-Arg in which theN-terminal amino acid is in the D form have-proven particularlybeneficial. D-Phe-Pro-Arg isopropyl ester is described as a competitivethrombin inhibitor (C. Mattson et al., Folia Haematol, 109 (1983)43-51).

Derivatization of the arginine at the C terminus to the aldehyde leadsto an enhancement of the inhibitory effect. Thus, a large number ofarginals able to bind the hydroxyl group of the “active” serine in ahemiacetal have been described (EP 185390, 479489, 526877, 542525; WO93/15756, 93/18060.

The thrombin-inhibitory activity of peptide ketones, fluorinated alkylketones and of keto esters, boric acid derivatives, phosphoric estersand α-keto carboxamides can likewise be explained by this serineinteraction (EP 118280, 195212, 362002, 364344, 410411, 471651, 589741,293881, 503203, 504064, 530167; WO 92/07869, 94/08941).

The peptide 4-amidinophenylglycinephosphonate diphenyl esters describedby J. Oleksyszyn et al. in J. Med. Chem. 37 (1994) 226-231 areirreversible thrombin inhibitors with inadequate selectivity in respectof other serine proteases.

DE 3 108 810, WO 93/11152 and EP 601 459 describe agmatine and hencearginine derivatives which are unable to interact with the active serinein serine proteases.

WO 94/29336, EP 0 601 459 and WO 95/23609 represent a furtherdevelopment in which the agmatine is replaced by an arylamidine residue.

EP 0 672 658 describes not only thrombin inhibitors which have attachedto them an agmatine or benzamidine residue, but also a thrombininhibitor having an amidinothiophene (Example 65).

Kininogenases are serine proteases which liberate vasoactive peptides,called kinins (bradykinin, kallidin and Met-Lys-bradykinin), fromkininogens. Kininogens are multifunctional proteins which occur incoagulation and inflammation cascade reactions. As inhibitors, theyprotect cells from damage by cysteine proteases (Müller Esterl, FEBSLett. 182 (1985) 310-314). Important kininogenases are plasmakallikrein, tissue kallikrein and mast cell tryptase.

Kinins like bradykinin and kallidin are vasoactive peptides whichinfluence a large number of biological processes. They play an essentialpart in inflammatory processes. By increasing vascular permeability,they lead to hypotension and edema. Furthermore, they are very potentpain-producing substances produced naturally in the body and have greatimportance as cellular mediators in the pathophysiology of asthma, ofallergic rhinitis and of arthritis (K. D. Bhoola, C. D. Figueroa, K.Worthy, Pharmacological Reviews 44 (1) (1992) 1-80).

Irrespective of the mechanisms underlying inflammatory processes, fluidcontaining all the protein systems in the circulating blood escapes fromblood vessels. This means that escape of plasma fluid from vessels isinvolved in diseases such as asthma, rhinitis and inflammatory internaldiseases. Moreover, mast cell tryptase is released particularly inallergic processes (Salomonsson et al., Am. Rev. Respir. Dis. 146 (1992)1535-1542).

The arginine chloromethyl ketones H-(D)-Pro-Phe-Arg-CH₂Cl andH-(D)-Phe-Phe-Arg-CH₂-Cl have been described by Kettner and Shaw asplasma kallikrein inhibitors (Biochem. 17 (1978) 4778-4784 and Meth.Enzym. 80 (1981) 826-842).

Various synthetic derivatives of benzamidines and benzylamines haveproven to be inhibitors of plasma kallikrein, with the benzamidineshaving a considerably stronger inhibitory effect (F. Markward, S.Drawert, P. Walsmann, Biochemical Pharmacology 23 (1974) 2247-2256).

PKSI-527, the hydrochloride ofN-(trans-4-aminomethylcyclohexylcarbonyl)-L-phenylalanine4-carboxymethylanilide, is also an effective inhibitor of thiskininogenase (Wanaka, Ohamoto et al., Thromb. Res., 57 (6) (1990)889-895).

The invention relates to compounds of the formula I

A—B—D—E—F  I

in which A, B, D, E and F have the following meanings:

A:

 where

m is 0, 1 or 2,

n is 0, 1 or 2,

R¹ is HOOC—, C₁₋₆-alkyl-OOC—, aryl-C₀₋₄-alkyl-OOC or —OH,

R² is H—, C₁₋₄-alkyl- or R¹—(CH₂)_(m)— and

R³ is H— or C₁₋₄-alkyl-,

B:

 where

R⁴ is H—, C₁₋₄-alkyl- or R¹—(CH₂)_(m)— (where R¹ and m have theabovementioned meanings),

p is 0 or 1,

R⁵ is H— or C₁₋₄-alkyl-,

R⁶ is H—, C₁₋₈-alkyl-, 2-thienyl-, 3-thienyl-, 3-indolyl-,4-imidazolyl-, 2-pyridyl-, 3-pyridyl-, 4-pyridyl-, phenyl- which maycarry up to three identical or different radicals from the group ofC₁₋₄-alkyl-, CF₃—, C₁₋₄-alkoxy-, HO—, BnO—, F— or Cl—, orC₃₋₈-cycloalkyl- which may carry up to four identical or differentC₁₋₄-alkyl radicals and/or where one or two C—C single bonds in the ringcan be replaced by a C═C double bond and/or a phenyl ring can be fusedon, C₇-C₁₂-bicycloalkyl- or C₁₀-tricycloalkyl- or

R⁴ and R⁶ together are an ethylene or propylene group,

R⁷ is H, C₁₋₈-alkyl-, phenyl- which may carry up to three identical ordifferent radicals from the group of C₁₋₄-alkyl-, CF₃—, C₁₋₄-alkoxy-, F—or Cl—, or C₃₋₈-cycloalkyl- which may carry up to four identical ordifferent C₁₋₄-alkyl radicals, and

R⁸ is H or C₁₋₄-alkyl,

D:

where R²⁰ is H, C₁₋₄-alkyl, Bn or BnO(CO)— and where the followingapplies:

if D is II, III or XI, then E has the following meaning:

where

a) in the event that X=S, O, NH or NR¹²,

Y is —CR¹³═, —CH═ and

Z is —CR¹⁴═

or

Y is —CR¹³═ and

Z is —CH═

or

b) in the event that X=NR¹²,

Y is —CH═ and

Z is —CH═

or

c) in the event that X=S, O or NH,

Y is —CR¹⁵═ and

Z is —N═

or

Y is —N═ and

Z is —CR¹⁵═

or

d) in the event that X=—NR¹²—,

Y is —N═ and

Z is —CR¹⁶═, —N═

or

Y is —CR¹⁶═ and

Z is —N═

and

R⁹ is H— or C₁₋₃-alkyl-,

R¹⁰ is H— or C₁₋₄-alkyl-,

R¹¹ is H— or C₁₋₄-alkyl-,

R¹² is CH₃— or C₂H₅—,

R¹³ is Cl—, CF₃— or C₁₋₄alkyl-,

R¹⁴ is Cl—, CF₃— or C₁₋₄-alkyl-,

R¹⁵ is CF₃— or C₁₋₄-alkyl-,

R¹⁶ is H—, CF₃— or C₁₋₄-alkyl- and

R²⁰ is as above,

or, if D is IV, VI, VII, VIII, IX or X, then E has the followingmeaning:

where

X is O, S or —NR¹⁷—

and

Y is —N═ and

Z is —CR¹⁶═ or —N═

or

Y is —CR¹⁶═ and

Z is —N═

or

Y is —CR¹⁸═ and

Z is —CR¹⁹═

and

R⁹, R¹⁰, R¹¹, R¹⁶ and R²⁰ are as above,

R¹⁷ is H, CH₃— or C₂H₅—,

R¹⁸ is H—, Cl—, CF₃— or C₁₋₄-alkyl-,

R¹⁹ is H—, Cl—, CF₃— or C₁₋₄-alkyl-,

or

if D is II, III, IV, VI, VII, VIII, IX, X or XI, E has the followingmeanings:

where

a) in the event that X=S,

Y is —CR¹⁸═ and

Z is —CR¹⁹═

or

Y is —CR¹⁶═ and

Z is —N═

or

b) in the event that X=O or —NR¹²—,

Y is —N═, —CR¹⁶═ and

Z is —N═, —CR¹⁸═

and

R⁹, R¹⁰, R¹¹, R¹², R¹⁶, R¹⁸, R¹⁹ and R²⁰ have the abovementionedmeanings,

F:

and their salts with physiologically acceptable acids.

The amino acid derivatives represented by B preferably have the (D)configuration; azetidinecarboxylic acid, proline and pipecolic acid in Dpreferably have the (L) configuration.

Preferred compounds of the formula I are those where A to E have thefollowing meanings:

A:

HOOC—(CH₂)_(t)— (t=1, 2 or 3), (HOOC—CH₂)₂—CH—,

HOOC—CH₂—CH(COOH)—, HOOC—CH(C₁₋₄-alkyl)-,

HOOC—C(C₁₋₄-alkyl)₂—, C₁₋₆-alkyl-OOC—(CH₂)_(t)—,

B is

p is 0 or 1,

R⁴ is H—, C₁₋₄-alkyl- or HOOC—(CH₂)_(m)— (m=1, 2 or 3),

R⁵ is H—, methyl-

R⁶ is H—, C₁₋₈-alkyl-, 2-thienyl-, 3-thienyl-, 3-indolyl-,4-imidazolyl-, 2-pyridyl-, 3-pyridyl-, 4-pyridyl-, phenyl- which maycarry up to three identical or different radicals from the group ofCH₃—, CF₃—, CH₃—O—, HO—, BnO—, F— or Cl—, or C₃₋₈-cycloalkyl, which maycarry up to four methyl radicals, bicyclo[2.2.2]octyl-,bicyclo[2.2.1]heptyl-, adamantyl-, indanyl-, decalinyl-,

R⁷ is H, C₁₋₈-alkyl-, phenyl-, which may carry up to three identical ordifferent radicals from the group of CH₃—, CF₃—, CH₃O—, F— or Cl—, orC₃₋₈-cycloalkyl- which may carry up to four methyl radicals,

R⁸ is H, C₁₋₄-alkyl,

D:

where R²⁰ is H, CH₃, Bn oder BnO(CO)— and

where the following applies:

if D is II, III or XI, then E has the meaning:

where

a) in the event that X=S, O or NR¹⁷,

Y is —CR¹³═ or —CH═ and

Z is —CR¹⁴═

or

Y is —CR¹³═ and

Z is —CH═

or

b) in the event that X=NR¹²,

Y is —CH═ and

Z is —H═,

or

c) in the event that X=S, O or NH,

Y is —CR¹⁵═ and

Z is —N═

or

Y is —N═ and

Z is —CR¹⁵═

or

d) in the event that X=NR¹²,

Y is —N═ and

Z is —CR¹⁶═, —N═

or

Y is —CR¹⁶═ and

Z is —N═

and

R¹² is CH₃— or C₂H₅—,

R¹³ is Cl—, CF₃— or C₁₋₄-alkyl-,

R¹⁴ is Cl—, CF₃— or C₁₋₄-alkyl-,

R¹⁵ is CF₃— or C₁₋₄alkyl-,

R¹⁶ is H—, CF₃— or C₁₋₄-alkyl-,

R¹⁷ is H, CH₃— or C₂H₅—

R²⁰ is as above, or

if D is IV, VI, VII, VIII, IX or X, then E has the meaning:

where

X is O, S or —NR¹⁷— and where

Y is —N═ and

Z is —CR¹⁶═ or —N═

or

Y is —CR¹⁶═ and

Z is —N═

or

Y is —CR¹⁸═ and

Z is —CR¹⁹═

and

R¹⁶, R¹⁷, R²⁰ have the abovementioned meanings,

R¹⁸ is H—, Cl—, CF₃— or C₁₋₄-alkyl- and

R¹⁹ is H—, Cl—, CF₃— or C₁₋₄-alkyl-,

or

if D is II, III, IV, VI, VII, VIII, IX, X or XI, then E has themeanings:

where

a) in the event that X=S,

Y is —CR¹⁸═ and

Z is —CR¹⁹═

or

Y is —CR¹⁶═ and

Z is —N═

or

b) in the event that X=O or —NR¹²—,

Y is —N═ or —CR¹⁶═ and

Z is —N═ or —CR¹⁸═

and R¹², R¹⁶, R¹⁸, R¹⁹ and R²⁰ have the abovementioned meanings,

F:

and their salts with physiologically acceptable acids.

The amino acid derivatives represented by B preferably have the (D)configuration; azetidinecarboxylic acid, proline and pipecolic acid in Dpreferably have the (L) configuration.

Especially preferred compounds of the formula I are those where A, B, D,E and F have the following meanings:

A: HOOC—CH₂, HOOC—CH₂—CH₂, HOOC—CH(CH₃), HOOC—CH(C₂H₅)

p is 0 or 1,

R⁴ is H—, CH₃—

R⁵ is H—, CH₃—,

R⁶ is C₁₋₈-alkyl-, C₅₋₈-cycloalkyl- which may carry up to four methylradicals, 2-thienyl-, 3-indolyl-, 4-imidazolyl-, 2-pyridyl-, 3-pyridyl-,4-pyridyl, phenyl- which may carry up to three identical or differentradicals from the group of CH₃—, CF₃—, CH₃O—, HO—, BnO—, F— or Cl—,bicyclo[2.2.2]octyl, bicyclo[2.2.1]heptyl, adamantyl, indanyl,decalinyl, with particular emphasis on cyclopentyl, cyclohexyl andcycloheptyl,

R⁷ is H, CH₃—,

R⁸ is H, CH₃—,

D:

where R²⁰ is H, BnO(CO)— and

where the following applies:

if D is II, III or XI, then E has the meaning

where

X is —S— and where

Y is —CH═ and

Z is —CR¹³═

or

Y is —CR¹³═ and

Z is —CH═

or

Y is —CR¹⁵═ and

Z is —N═

or

Y is —N═ and

Z is —CR¹⁵═

and

R¹³ is Cl—, CF₃— or CH₃—

R¹⁵ is CF₃— or CH₃— and

R²⁰ is as above,

or

if D is IV, VI, VII, VIII, IX or X, then E has the meaning:

where

X is S and where

Y is —N═ and

Z is —CR¹⁶═

or

Y is —CR¹⁶═ and

Z is —N═

or

Y is —CR¹³═ and

Z is —CH═

or

Y is —CH═ and

Z is —CR¹³═

or

Y is —CH═ and

Z is —CH═

and

R¹³, R²⁰ have the abovementioned meanings and

R¹⁶ is H—, CF₃— or CH₃—

or

if D is II, III, IV, VI, VII, VIII, IX, X or XI, then E has themeanings:

where either

a) in the event that X=S,

Y is —CH═ and

Z is —CR¹⁸═

or

Y is —CR¹⁶═ and

Z is —N═

or

Y is —CR¹⁸═ and

Z is —CH═

or

b) in the event that X=O or NCH₃

Y is —CH═ and

Z is —CR¹⁶═

or

Y is —CR¹⁶═ and

Z is —CH═

or

c) in the event that X=—NR¹²—

Y is —N═ and

Z is —CR¹⁸═

and

R¹² is CH₃— or C₂H₅— and

R¹⁸ is H, Cl—, CF₃— or CH₃—, and

R¹⁶, R²⁰ have the abovementioned meanings

F:

and their salts with physiologically acceptable acids.

The amino acid derivatives represented by B preferably have the (D)configuration; azetidinecarboxylic acid, proline and pipecolic acid in Dpreferably have the (L) configuration.

Very especially preferred compounds of the formula I are those where A,B, D, E and F have the following meanings:

A: HOOC—CH₂, HOOC—CH₂—CH₂, HOOC—CH(CH₃), HOOC—CH(C₂H₅)

B:

p is 0 or 1,

R⁴ is H—,

R⁵ is H—,

R⁶ is C₁₋₈-alkyl-, 2-thienyl-, 3-indolyl-, 4-imidazolyl-, 2-pyridyl-,3-pyridyl-, 4-pyridyl-, C₅₋₈-cycloalkyl- which may carry up to fourmethyl radicals, phenyl- which may carry up to three identical ordifferent radicals from the group of CH₃—, CF₃—, CH₃O—, HO—, BnO—, F— orCl—, bicyclo[2.2.2]octyl, bicyclo[2.2.1]heptyl, adamantyl, indanyl,decalinyl, with particular emphasis on cyclopentyl-, cyclohexyl- andcycloheptyl-,

R⁷ is H,

R⁸ is H,

D:

where the following applies:

if D is II, III or XI, then E has the meaning

where

X is S and

Y is —CR¹³═ and

Z is —CH═

or

Y is —CH═ and

Z is —CR¹³═

or

Y is —CR¹⁵═ and

Z is —N═

or

Y is —N═ and

Z is —CR¹⁵═

and

R¹³ is Cl—, CF₃— or CH₃— and

R¹⁵ is CF₃— or CH₃—,

or

if D is IV, VI, VII, VIII, IX or X, then E has the meaning

where

X is S and

Y is —N═ and

Z is —CR¹⁶═

or

Y is —CR¹⁶═ and

Z is —N═

or

Y is —CH═ and

Z is —CR¹³═

or

Y is —CR¹³═ and

Z is —CH═

or

Y is —CH═ and

Z is —CH═

and

R¹³ has the abovementioned meaning and

R¹⁶ is H, CF₃— or CH₃—, or

if D is II, III, IV, VI, VII, VIII, IX, X or XI, then E has the meanings

where

a) in the event that X=S,

Y is —CH═ and

Z is —CR¹⁸═

or

Y is —CR¹⁸═ and

Z is —CH═

or

Y is —CR¹⁶═ and

Z is —N═

or

b) in the event that X=O or NCH₃

Y is —CH═ and

Z is —CR¹⁶═

or

Y is —CR¹⁶═ and

Z is —CH═

or

c) in the event that X=NCH₃

Y is —N═ and

Z is —CR¹⁶═

and

R¹⁶ has the abovementioned meaning and

R¹⁸ is H, Cl— CF₃— or CH₃—,

F:

and their salts with physiologically acceptable acids.

The amino acid derivatives represented by B preferably have the (D)configuration; azetidinecarboxylic acid, proline and pipecolic acid in Dpreferably have the (L) configuration.

With the exception of the compounds mentioned in the Examples, thefollowing substances must be very especially emphasized:

HOOC—CH₂-(D)-Cha-Pro-NH—CH₂-5-(2-am-3-CF₃)-thioph

HOOC—CH₂-(D)-Chg-Pro-NH—CH₂-5-(2-am-3-CF₃)-thioph

HOOC—CH₂-(D)-Cha-Pro-NH—CH₂-5-(2-am-4-Me)-thioph

HOOC—CH₂-(D)-Cha-Pro-NH—CH₂-5-(2-am-4-Cl)-thioph

HOOC—CH₂-(D)-Cha-Pro-NH—CH₂-5-(2-am-4-CF₃)-thioph

HOOC—CH₂-(D)-Chg-Pro-NH—CH₂-5-(2-am-4-Me)-thioph

HOOC—CH₂-(D)-Chg-Pro-NH—CH₂-5-(2-am-4-Cl)-thioph

HOOC—CH₂-(D)-Chg-Pro-NH—CH₂-5-(2-am-4-CF₃)-thioph

HOOC—CH₂-(D)-Chg-Pro-NH—CH₂-5-(2-am-3,4-Me₂)-thioph

HOOC—CH₂-(D)-Cha-Aze-NH—CH₂-2-(4-am)-thioph

HOOC—CR₂-(D)-Chg-Pic-NH—CH₂-2-(5-am)-thioph

HOOC—CH₂-(D)-Cha-Pic-NH—CH₂-2-(4-am)-thioph

HOOC—CH₂-(D)-Chg-Pic-NH—CH₂-2-(4-am)-thioph

HOOC—CH₂-(D)-Cha-Pic-NH—CH₂-2-(5-am-3-Me)-thioph

HOOC—CH₂-(D)-Chg-Pic-NH—CH₂-2-(5-am-3-Me)-thioph

HOOC—CH₂-(D)-Cha-Pic-NH—CH₂-2-(5-am-3-Cl)-thioph

HOOC—CH₂-(D)-Chg-Pic-NH—CH₂-2-(5-am-3-Cl)-thioph

HOOC—CH₂-(D)-Cha-Pic-NH—CH₂-2-(5-am-4-Me)-thioph

HOOC—CH₂-(D)-Chg-Pic-NH—CH₂-2-(5-am-4-Me)-thioph

HOOC—CH₂-(D)-Cha-Pic-NH—CH₂-2-(5-am-4-Cl)-thioph

HOOC—CH₂-(D)-Chg-Pic-NH—CH₂-2-(5-am-4-Cl)-thioph

HOOC—CH₂-(D)-Chea-Pro-NH—CH₂-2-(4-am)-thioph

HOOC—CH₂-(D)-Cpa-Pro-NH—CH₂-2-(4-am)-thioph

HOOC—CH₂-(D)-Chg-Pro-NH—CH₂-2-(4-am-5-Me)-thioph

HOOC—CH₂-(D)-Chg-Pro-NH—CH₂-2-(4-am-5-Cl)-thioph

HOOC—CH₂-(D)-Chg-Pro-NH—CH₂-2-(4-am-5-CF₃)-thioph

HOOC—CH₂-CH₂-(D)-Chg-Pro-NH—CH₂-2-(4-am)-thioph

HOOC—CH₂-CH₂-(D)-Cha-Pro-NH—CH₂-2-(4-am)-thioph

HOOC—CH₂-(D)-Chea-Pro-NH—CH₂-4-(2-am)-thioph

HOOC—CH₂-(D)-Cpa-Pro-NH—CH₂-4-(2-am)-thioph

HOOC—CH₂-(D)-Chg-Pro-NH—CH₂-4-(2-am)-thioph

HOOC—CH₂-(D)-Cheg-Pro-NH—CH₂-4-(2-am)-thioph

HOOC—CH₂-(D)-Cpg-Pro-NH—CH₂-4-(2-am)-thioph

HOOC—CH₂-(D)-Cha-Pic-NH—CH₂-2-(4-am)-thiaz

HOOC—CH₂-(D)-Chg-Pic-NH—CH₂-2-(4-am)-thiaz

HOOC—CH₂-(D)-Cha-Aze-NH—CH₂-2-(4-am)-thiaz

HOOC—CH₂-(D)-Chg-Aze-NH—CH₂-2-(4-am)-thiaz

MeOOC—CH₂-(D)-Cha-Pro-NH—CH₂-2-(4-am)-thiaz

HOOC—CH₂-(D)-Cpg-Pro-NH—CH₂-2-(4-am)-thiaz

HOOC—CH₂-(D)-Chea-Pro-NH—CH₂-2-(4-am)-thiaz

HOOC—CH₂-(D)-Cheg-Pro-NH—CH₂-2-(4-am)-thiaz

HOOC—CH₂-(D)-Cha-Pic-NH—CH₂-2-(4-am)-thiaz

HOOC—CH₂-(D)-Chg-Pic-NH—CH₂-2-(4-am)-thiaz

HOOC—CH₂-CH₂-(D)-Cha-Pro-NH—CH₂-2-(4-am)-thiaz

HOOC—CH₂-CH₂-(D)-Chg-Pro-NH—CH₂-2-(4-am)-thiaz

HOOC—CH₂-(D)-Cha-Pro-NH—CH₂-2-(4-am-5-Me)-thiaz

HOOC—CH₂-(D)-Chg-Pro-NH—CH₂-2-(4-am-5-Me)-thiaz

HOOC—CH₂-(D)-Cha-Pro-NH—CH₂-2-(4-am-5-CF₃)-thiaz

HOOC—CH₂-(D)-Chg-Pro-NH—CH₂-2-(4-am-5-CF₃)-thiaz

HOOC—CH₂-(D)-Cha-Pro-NH—CH₂-2-(5-am-4-Me)-thiaz

HOOC—CH₂-(D)-Chg-Pro-NH—CH₂-2-(5-am-4-Me)-thiaz

HOOC—CH₂-(D)-Cha-Pro-NH—CH₂-2-(5-am-4-CF₃)-thiaz

HOOC—CH₂-(D)-Chg-Pro-NH—CH₂-2-(5-am-4-CF₃)-thiaz

HOOC—CH₂-(D)-Chg-Pro-NH—CH₂-5-(3-am)-isox

HOOC—CH₂-(D)-Cha-Pro-NH—CH₂-2-(4-am)-oxaz

HOOC—CH₂-(D)-Chg-Pro-NH—CH₂-2-(4-am)-oxaz

HOOC—CH₂-CH₂-(D)-Chg-Pro-NH—CH₂-5-(3-am)-fur

HOOC—CH₂-CH₂-(D)-Cha-Pro-NH—CH₂-5-(3-am)-fur

HOOC—CH₂-(D)-Chea-Pro-NH—CH₂-5-(3-am)-fur

HOOC—CH₂-(D)-Cha-Pic-NH—CH₂-5-(3-am)-fur

HOOC—CH₂-(D)-Chg-Pic-NH—CH₂-5-(3-am)-fur

HOOC—CH₂-CH₂-(D)-Cha-Pic-NH—CH₂-5-(3-am)-fur

HOOC—CH₂-CH₂-(D)-Chg-Pic-NH—CH₂-5-(3-am)-fur

HOOC—CH₂-(D)-Chg-Aze-NH—CH₂-5-(3-am)-fur

HOOC—CH₂-(D)-Cha-Aze-NH—CH₂-5-(3-am)-fur

HOOC—CH₂-(D)-Cha-Aze-NH—CH₂-2-(4-am-1-Me)-pyrr

HOOC—CH₂-(D)-Chg-Aze-NH—CH₂-2-(4-am-1-Me)-pyrr

HOOC—CH₂-(D)-Chg-Pro-NH—CH₂-2-(4-am-1-Me)-pyrr

HOOC—CH₂-(D)-Cha-Pic-NH—CH₂-2-(4-am-1-Me)-pyrr

HOOC—CH₂-(D)-Chg-Pic-NH—CH₂-2-(4-am-1-Me)-pyrr

HOOC—CH₂-(D)-Cha-Aze-NH—CH₂-2-(5-am-1-Me)-pyrr

HOOC—CH₂-(D)-Chg-Aze-NH—CH₂-2-(5-am-1-Me)-pyrr

HOOC—CH₂-(D)-Chg-Pro-NH—CH₂-2-(5-am-1-Me)-pyrr

HOOC—CH₂-(D)-Cha-Pic-NH—CH₂-2-(5-am-1-Me)-pyrr

HOOC—CH₂-(D)-Chg-Pic-NH—CH₂-2-(5-am-1-Me)-pyrr

HOOC—CH₂-(D)-Cha-Aze-NH—CH₂-4-(2-am-1-Me)-pyrr

HOOC—CH₂-(D)-Chg-Aze-NH—CH₂-4-(2-am-1-Me)-pyrr

HOOC—CH₂-(D)-Chg-Pro-NH—CH₂-4-(2-am-1-Me)-pyrr

HOOC—CH₂-(D)-Cha-Pic-NH—CH₂-4-(2-am-1-Me)-pyrr

HOOC—CH₂-(D)-Chg-Pic-NH—CH₂-4-(2-am-1-Me)-pyrr

HOOC—CH₂-(D)-Cha-Aze-NH—CH₂-5-(3-am-1-Me)-pyraz

HOOC—CH₂-(D)-Chg-Aze-NH—CH₂-5-(3-am-1-Me)-pyraz

HOOC—CH₂-(D)-Chg-Pro-NH—CH₂-5-(3-am-1-Me)-pyraz

HOOC—CH₂-(D)-Cha-Pic-NH—CH₂-5-(3-am-1-Me)-pyraz

HOOC—CH₂-(D)-Chg-Pic-NH—CH₂-5-(3-am-1-Me)-pyraz

HOOC—CH₂-(D)-Cha-Aze-NH—CH₂-3-(5-am-1-Me)-pyraz

HOOC—CH₂-(D)-Chg-Aze-NH—CH₂-3-(5-am-1-Me)-pyraz

HOOC—CH₂-(D)-Cha-Pro-NH—CH₂-3-(5-am-1-Me)-pyraz

HOOC—CH₂-(D)-Chg-Pro-NH—CH₂-3-(5-am-1-Me)-pyraz

HOOC—CH₂-(D)-Cha-Pic-NH—CH₂-3-(5-am-1-Me)-pyraz

HOOC—CH₂-(D)-Chg-Pic-NH—CH₂-3-(5-am-1-Me)-pyraz

HOOC—CH₂-(D)-Chg-Pro-NH—CH₂-5-(3-am)-oxadiaz

tBuOOC—H₂-N-BOC-(D)-Chg-Pro-NH—CH₂-2-(4-am)-oxaz

HOOC—CH₂-(D)-Chg-Pro-NH—CH₂-5-(3-am-4-Cl)-thioph

EtOOC—CH₂-(D)-Chg-Pro-NH—CH₂-5-(3am-4-Cl)-thioph

HOOC—CH₂-(D)-Chg-Pro-NH-H₂-5-(3-am-4-Me)-thioph

EtOOC—CH₂-(D)-Chg-Pro-NH—CH₂-5-3-am-4-Me)-thioph

tBuOOC—CH₂-(D)-Cha-Pro-NH—CH₂-4-(2-ham)-thioph

tBuOOC—CH₂-(D)-Chg-Pro-NH—CH₂-4-(2-ham)-thioph

tBuOOC—H₂-(D)-Chg-Aze-NH—CH₂-4-(2-ham)-thioph

tBuOOC—CH₂-(D)-Cha-Aze-NH—CH₂-4-(2-ham)-thioph

tBuOOCCH₂-(D)-Cha-Pro-NH—CH₂-4-(2-ham)-thiaz

tBuOOC—CH₂-(D)-Chg-Pro-NH—CH₂-4-(2-ham)-thiaz

tBuOOC—CH₂-(D)-Chg-Aze-NH—CH₂-4-(2-ham)-thiaz

tBuOOC—CH₂-(D)-Cha-Aze-NH—CH₂-4-(2-ham)-thiaz

List of abbreviations: Adaala: adamantylalanine Adagly: adamantylglycineAIBN: azobisisobutyronitrile Ac: acetyl am: amidino Aze:azetidinecarboxylic acid Bn: benzyl bs: broad singulet Boc:tert-butyloxycarbonyl Bu: butyl Cbz: benzyloxycarbonyl Cha:cyclohexylalanine Chea: cycloheptylalanine Cheg: cycloheptylglycine Chg:cyclohexylglycine Cog: cyclooctylglycine Cpa: cyclopentylalanine Cpg:cyclopentylglycine d: doublet TLC: Thin-layer chromatography DCC:dicyclohexylcarbodiimide Dch: dicyclohexylalanine Dcha:dicyclohexylamine DCM: dichloromethane Dep: 4,5-dehydropipecolic acidDMF: dimethylformamide DIPEA: diisopropylethylamine Dpa: diphenylalanineEDC: N-ethyl-N′-(3-dimethylaminopropyl)carbodiimide- hydrochloride Et:ethyl Eq: equivalents Gly: glycine fur: furan ham: hydroxyamidinoHOSucc: hydroxysuccinimide HPLC: high-performance liquid chromatographyimi: imidazole iPr: isopropyl isox: isoxazole Leu: leucine Lsg: solutionMe: methyl α-MeCha: α-methylcyclohexylalanine β, β-Me₂Cha:2-amino-3-cyclohexyl-3-methylbutyric acid orβ,β-dimethylcyclohexylalanine 4-MeCha: (4-methylcyclohex-1-yl)alanineγ-MeCha: (1-methylcyclohex-1-yl)alanine 3,3-Me₂Cha:(3,3-dimethylcyclohex-1-yl)alanine 4-MeChg: (4-methylcyhex-1-yl)glycine[sic] 3,3-Me₂Chg: (3,3-dimethylcyclohex-1-yl)glycine MPLC:medium-pressure liquid chromatography MTBE: methyl tert-butyl ether NBS:N-bromosuccinimide Nog: norbornylglycine Ohind:(2)-octahydroindole-2-carboxylic acid Oxadiaz: 1,2,4-oxadiazole Oxaz:oxazole Ph: phenyl Phe: phenylalanine Pic: pipecolic acid PPA:propylphosphonic anhydride Pro: proline Py: pyridine pydaz:(35)-2,3,4,5-tetrahydropyridazine-3-carboxylic acid Pyr:3,4-dehydroproline pyraz: pyrazole pyrr: pyrrole pyzo-3:(3S)pyrazolidine-3-carboxylic acid q: quartet RT: room temperatureRP-18: reversed phase C-18 s: singulet sbr: singulet, broad t: triplett: tertiary tBu: tertiary-butyl tert: tertiary TBAB: tetrabutylammoniumbromide TEA: trietylamine [sic] TFA: trifluoroacetic acid TFFA:trifluoroacetic anhydride thiaz: thiazole thioph: thiophene Thz-2:thiazolodine-2-carboxylic [sic] acid Thz-4: thiazolidine-4-carboxylicacid 5,5-Me₂Thz-4: (45)-5.5-dimethylthiazolidine-4-carboxylic acid TOTU:O-(cyanoethoxycarbonylmethylene)amino- N,N,N′,N′-tetramethyluroniumtetrafluoroborate triaz: 1,2,4-triazole Z: benzyloxycarbonyl

In the description and the claims, the following definitions apply tothe individual substituents:

The term “cycloalkyl”, on its own or as part of another substituentcomprises saturated or cyclic hydrocarbon groups which contain the givennumber of carbon atoms. C₃₋₈-cycloalkyl refers to saturated alicyclicrings having 3 to 8 C atoms such as, for example, cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, 4-methylcyclohexyl, cycloheptyl orcyclooctyl.

The term “alkyl” on its own or as part of another substituent denotes alinear or branched alkyl chain radical of the length indicated in eachcase. Thus, C₁₋₄-alkyl is, for example, methyl, ethyl, 1-propyl,2-propyl, 2-methyl-2-propyl, 2-methyl-1-propyl, 1-butyl, 2-butyl,C₁₋₆-alkyl, for example C₁₋₄-alkyl, pentyl, 1-pentyl, 2-pentyl,3-pentyl, 1-hexyl, 2-hexyl, 3-hexyl, 4-methyl-1-pentyl or3,3-dimethylbutyl. In addition to the radicals given for C₁₋₄-alkyl,C₁₋₈-alkyl denotes, for example, C₁₋₆-alkyl, heptyl or octyl.

The term “alkoxy” on its own or as part of another substituent denotes alinear or branched alkyl chain radical which has the length indicated ineach case and which is bonded to the respective basic compound via anoxygen atom. Thus C₁₋₄-alkoxy denotes, for example, methoxy, ethoxy,1-propoxy, 2-propoxy, 2-methyl-2-propoxy, 2-methyl-1-propoxy, 1-butoxy,2-butoxy.

The invention furthermore relates to compounds which contain thestructural element

where D and E have the abovementioned meanings and where a hydrogenatom, a protective group, an unsubstituted or substituted natural orunnatural amino acid, an unsubstituted or substituted carboxylic acid oran unsubstituted or substituted alkyl radical is located on the nitrogenatom of building block D. The structural fragment is valuable as acomponent of serine protease inhibitors and, in particular, of thrombinand kallikrein inhibitors.

The invention also relates to compounds which contain the structuralelement

where E has the abovementioned meaning and where a hydrogen atom, aprotective group, an unsubstituted or substituted natural or unnaturalamino acid, an unsubstituted or substituted carboxylic acid or anunsubstituted or substituted alkyl radical is located on the nitrogenatom of NR⁹.

Finally, the invention also relates to compounds which have one of thefollowing structural elements:

where Q is CH₃ or Cl; T is NCH₃, O or S; and W is NCH₃ or S.

The invention furthermore relates to the intermediates of the formulaeVa and Vb

A—B—D—E—CN  Va,

A—B—D—E—CSNH₂  Vb,

where A, B, D and E have the abovementioned meanings.

The novel intermediates are used to prepare the compounds I and arevaluable building blocks for synthesizing serine protease inhibitors.

The compounds of the formula I can exist as such or in the form of theirsalts with physiologically acceptable acids. Examples of such acids are:hydrochloric acid, citric acid, tartaric acid, lactic acid, phosphoricacid, methanesulfonic acid, acetic acid, formic acid, maleic acid,fumaric acid, succinic acid, hydroxysuccinic acid, sulfuric acid,glutaric acid, aspartic acid, pyruvic acid, benzoic acid, glucuronicacid, oxalic acid, ascorbic acid and acetylglycine.

If, in the compounds of the formula I, R¹ equals C₁₋₆-alkyl-OOC,aryl-C₀₋₄-alkyl-OOC and/or F equals hydroxyamidine, these compounds mayact in vivo as prodrugs from which the corresponding carboxylic acidsR¹═HOOC— or the corresponding amidines F═—C(═NH)-NH₂ are formedenzymatically.

Prodrugs of the compounds of the formula I are to be understood asmeaning those compounds which are metabolized in vivo to give thepharmacologically active compounds of the formula I. This can beeffected, for example, by the first-pass metabolism in the liver.

The novel compounds of the formula I are competitive inhibitors oftrypsin-like serine proteases, especially of thrombin, and also ofkininogenases such as kallikrein. They can be employed for the followingindications:

diseases whose pathogenetic mechanism derives directly or indirectlyfrom the proteolytic effect of thrombin,

diseases whose pathogenetic mechanism derives from thrombin-dependentactivation of receptors and signal transductions,

diseases associated with stimulation [eg. by PAI-1, PDGF (plateletderived growth factor), P-selectin, ICAM-1, tissue factor] or inhibition(eg. NO synthesis in smooth muscle cells) of the expression of genes inbody cells,

diseases deriving from the mitogenic effect of thrombin,

diseases deriving from a thrombin-dependent change in the contractilityand permeability of epithelial cells (eg. vascular endothelial cells),

thrombin-dependent thromboembolic events such as deep vein thrombosis,pulmonary embolism, myocardial or cerebral infarct, atrial fibrillation,bypass occlusion,

disseminated intravascular coagulation (DIC),

reocclusion and for reducing the reperfusion time on comedication withthrombolytics such as streptokinase, urokinase, prourokinase, t-PA,APSAC, plasminogen activators from the salivary glands of animals, andthe recombinant and mutated forms of all these substances,

the occurrence of early reocclusion and late restenosis after PTCA,

the thrombin-dependent proliferation of smooth muscle cells,

the accumulation of active thrombin in the CNS (eg. in Alzheimer'sdisease),

tumor growth and to prevent adhesion and metastasis of tumor cells.

The novel compounds can be used in particular for the therapy andprophylaxis of thrombin-dependent thromboembolic events such as deepvein thromboses, pulmonary embolisms, myocardial or cerebral infarctsand unstable angina, also for the therapy of disseminated intravascularcoagulation (DIC). They are furthermore suitable for combination therapywith thrombolytics such as streptokinase, urokinase, prourokinase, t-PA,APSAC and other plasminogen activators to shorten the reperfusion timeand extend the reocclusion time.

Further preferred areas of use are to prevent thrombin-dependent earlyreocclusion and late restenosis after percutaneous transluminal coronaryangioplasty, to prevent thrombin-induced proliferation of smooth musclecells, to prevent accumulation of active thrombin in the CNS (eg. inAlzheimer's disease), to control tumors and to prevent mechanisms whichlead to adhesion and metastasis of tumor cells.

The novel compounds can also be used for coating artificial surfacessuch as hemodialysis membranes and the tubing systems and linesnecessary therefor, and for coating oxygenators in extravascularcirculation, stents and heart valves.

The novel compounds can furthermore be employed for diseases whosepathogenetic mechanism derives directly or indirectly from theproteolytic effect of kininogenases, especially kallikrein, eg. ininflammatory diseases such as asthma, pancreatitis, rhinitis, arthritis,urticaria and other internal inflammatory diseases.

The compounds according to the invention can be administered in aconventional way orally or parenterally (subcutaneously, intravenously,intramuscularly, interperitoneally, rectally). Administration can alsotake place with vapors or sprays through the nasopharyngeal space.

The dosage depends on the age, condition and weight of the patient andon the mode of administration. As a rule, the daily dose of activesubstance per person is about 10-2000 mg on oral administration andabout 1-200 mg on parenteral administration. This dose can be given in 2to 4 single doses or once a day as depot form.

The novel compounds can be used in conventional solid or liquidpharmaceutical forms, eg. as uncoated or (film-)coated tablets,capsules, powders, granules, sugar-coated tablets, suppositories,solutions, ointments, creams or sprays. These are produced in aconventional manner. The active substances can for this purpose be mixedwith conventional pharmaceutical auxiliaries such as tablet binders,bulking agents, preservatives, tablet disintegrants, flow regulators,plasticizers, wetting agents, dispersants, emulsifiers, solvents,release-slowing agents, antioxidants and/or propellant gases (cf. H.Sucker et al.: Pharmazeutische Technologie, Thieme-Verlag, Stuttgart,1978). The administration forms obtained in this way normally containfrom 0.1 to 99% by weight of active substance.

EXPERIMENTAL PART

The compounds of the formula I can be prepared as shown in SchemesI-III.

Building blocks A, B, D and E are preferably assembled separatelybeforehand and employed in suitably protected form (see Scheme I-III).

Scheme I describes the linear assemblage of the molecule I by couplingthe amine H—E—CN to the N-protected amino acid P—D—OH to give P—D—E—CN,eliminating the N-terminal protective group to give H—D—E—CN, couplingto the N-protected amino acid P—B—OH to give P—B—D—E—CN, eliminating theprotective group P to give H—B—D—E—CN, subsequently alkylating with theunprotected or protected (P)—A—U building block (U=leaving group) orreductively alkylating with (P)—A′—U (U=aldehyde, ketone) or Michaeladdition with a suitable (P)—A″—C═C— derivative to give (P)—A—B—D—E—CN.Conversion of the nitrile functionality into the amidine group takesplace either by the classical Pinner synthesis (R. Boder, D. G. Neilson,Chem. Rev. 61 (1962) 179) or by a modified Pinner synthesis whichproceeds via imino thioester salts as intermediate (H. Vieweg et al.,Pharmazie 39 (1984) 226) or directly by the method of A. EschenmoserHelv. Chimica Acta 69 (1986) 1224. Subsequently the protective groupsstill present in the molecule are eliminated, preferably by acidhydrolysis.

If building block E is incorporated as H—E—CONH₂ into the synthesis,dehydration of the amide to the.nitrile functionality or conversion tothe thioamide functionality takes place on one of the protectedintermediates. As an alternative, the building block E may be employedin the synthesis in the form of H—E—CSNH₂.

Scheme II describes the linear assemblage of the molecule I byalkylation, reductive amination or Michael addition of H—B—P ontoappropriately suitable unprotected or protected A building blocks togive (P)—A—B—P, elimination of the C-terminal protective group to give(P)—A—B—OH, coupling to H—D—P to give (P)—A—B—D—P, elimination of theC-terminal protective group to give (P)—A—B—D—OH, coupling to H—E—CN togive (P)—A—B—D—E—CN and reaction of this intermediate to give the finalproduct as in Scheme I.

Where compounds (P)—A—B—P still have a free NH functionality on B, thismust be provided with a suitable protective group before elimination ofthe C-terminal protective group. The protective groups used in each casemust be orthogonal to one another.

As an alternative to the H—E—CN building block, it is also possible toemploy H—E—CONH₂, H—E—CSNH₂, H—E—C(NH)NH₂, H—E—C(NP)NH₂, H—E—C(NP)NHP,with the coupled intermediate (P)—A—B—D—E—CONH₂ in the first case beingdehydrated to (P)—A—B—D—E—CN or being converted directly into(P)—A—B—D—E—CSNH₂, for example by means of Lawesson's reagent.

Scheme III describes a very efficient way for preparing compounds I by aconvergent synthesis. The appropriately protected building blocks(P)—A—B—OH and H—D—E—CN are coupled together and the resultingintermediate (P)—A—B—D—E—CN is reacted to give the final product as inScheme I.

It is also possible to employ H—D—E—CONH₂ or H—D—E—CSNH₂ as analternative to H—D—E—CN, with the coupled intermediate (P)—A—B—D—E—CONH₂in the first case being dehydrated to (P)—A—B—D—E—CN or being convertedinto (P)—A—B—D—E—CSNH₂.

The N-terminal protective groups employed are Boc, Cbz or Fmoc,preferably Boc, and the C-terminal protective groups are methyl,tert-butyl and benzyl. If a plurality of protective groups is present inthe molecule, they must be orthogonal to one another if they are not tobe eliminated simultaneously.

The required coupling reactions and the other reactions for introducingand eliminating protective groups are carried out under standardconditions of peptide chemistry (see M. Bodanszky, A. Bodanszky “ThePractice of Peptide Synthesis”, 2nd edition, Springer Verlag Heidelberg,1994).

Boc protective groups are eliminated using dioxane/HCl or TFA/DCM, andCbz protective groups are eliminated by hydrogenolysis or with HF.Hydrolysis of ester functionalities takes place with LiOH in analcoholic solvent or in dioxane/water. TFA or HCl are used to cleavet-butyl esters.

The reactions were checked by TLC, normally using the following mobilephases:

A. DCM/MeOH 95:5 B. DCM/MeOH  9:1 C. DCM/MeOH  8:2 D. DCM/MeOH/50% HOAc40:10:5 E. DCM/MeOH/50% HOAc 35:15:5

Where separations by column chromatography are mentioned, these wereseparations on silica gel using the abovementioned mobile phases.

Reversed phase HPLC separations were carried out withaceto-nitrile/water and HOAc buffer.

All reactions were routinely carried out under a nitrogen atmosphere.

The starting compounds can be prepared by the following methods:

Examples of building blocks A employed for the alkylation are tert-butylα-bromoacetate, tert-butyl β-bromopropionate, tert-butylα-bromopropionate, tert-butyl γ-bromobutyrate, tert-butylα-bromobutyrate, THP-protected bromoethanol, THP-protectedγ-bromopropanol, α-bromo-γ-butyrolactone, for the reductive aminationare dihydroxyacetone, di-tert-butyl acetonedicarboxylate, and for theMichael addition are tert-butyl acrylate, tert-butyl methacrylate,di-tert-butyl fumarate. Those of said tert-butyl esters which cannot bepurchased are prepared by methods similar to G. Uray, W. Lindner,Tetrahedron, 44 1988 357-4362.

B building blocks:

A wide variety of possibilities is available in the literature for thegeneral and specific synthesis of amino acids. A review thereof isprovided by, inter alia, Houben-Weyl, Volume E16d/Part 1, pages 406 etseq.

Precursors which were frequently employed were benzophenone imine aceticacid ethyl ester, diethyl-acetamidomalonate and ethyl isonitrileacetate.

Various racemic glycine and alanine derivatives were prepared, forexample, starting from ethyl isonitrileacetate and an appropriate ketoneor aldehyde (see H.-J. Prätorius, J. Flossdorf, M.-R. Kula Chem. Ber.108 (1975) 3079).

The syntheses of cyclooctylglycine, cycloheptylglycine,2-norbonylglycine [sic], adamantylalanine, γ-methylcyclohexylalanine,4-isopropyl-1-cyclohexylalanine, 4-methyl-1-cyclohexylalanine,4-methyl-1-cyclohexylglycine, cycloheptylalanine and cyclopentyl-alaninewere carried out via the corresponding ethyl 2-formylaminoacrylates (U.Schöllkopf and R. Meyer, Liebigs Ann. Chem. 1977, 1174 and H.-J.Prätorius, J. Flossdorf, M.-R. Kula Chem. Ber. 108 (1985) 3079) startingfrom ethyl isocyanoacetate with the relevant carbonyl compoundscyclooctanone, cyclo-heptanone, 2-norbornanone, 1-formyladamantane,1-formyl-1-methylcyclohexane, 1-formyl-4-isopropylcyclohexane,1-formyl-4-methylcyclohexane and 4-methyl-cyclohexanone,formylcyclohexane and formylcyclopentane by the following generalmethods:

General Method for Synthesizing Ethyl 2-Formylaminoacrylates

A solution of 100 m [sic] of ethyl isocyanoacetate in 50 ml of THF wasadded dropwise to 100 m [sic] of potassium tert-butoxide in 150 ml ofTHF at 0 to −10° C. After 15 min at the same temperature 100 mmol of theappropriate carbonyl compound in 50 ml of THF were added, the reactionmixture was allowed to rise slowly to RT, and the solvent was strippedoff in a rotary evaporator. The residue was mixed with 50 ml of water,100 ml of acetic acid and 100 ml of DCM, and the product was extractedwith DCM. The DCM phase was dried over Na₂SO₄, and the solvent isstripped off in a rotary evaporator. The resulting products were almostpure but could, if necessary, be purified further by columnchromatography on silica gel (mobile phases: ether/petroleum ethermixtures).

General Method for Amino Acid Hydrochlorides Starting From the Ethyl2-Formylaminoacrylates

100 m [sic] of the ethyl 2-formylaminoacrylates were hydrogenated withPd/C (10%) and hydrogen in 200 ml of glacial acetic acid until thereaction was complete. The catalyst was then filtered off, the aceticacid was stripped off as far as possible in a rotary evaporator, and theresidue was refluxed in 200 ml of 50% concentrated hydrochloric acid for5 h. The hydrochloric acid was stripped off in a rotary evaporator, andthe product was dried at 50° C. under reduced pressure and then washedseveral times with ether. The hydrochlorides resulted as pale coloredcrystals.

25.0 g of cyclooctylglycine hydrochloride were obtained starting from18.9 g (150 mmol) of cyclooctanone. 36.2 g of cycloheptylglycinehydrochloride were obtained starting from 22.4 g (200 mmol) ofcycloheptanone. 26.6 g of 2-norbonylglycine [sic] hydrochloride wereobtained starting from 16.5 g (150 mmol) of 2-norbornanone. 26.0 g ofadamantylalanine hydrochloride were obtained starting from 19.7 g (120mmol) of 1-formyladamantane. 16.6 g of y-methylcyclohexylalaninehydrochloride were obtained starting from 12.6 g (100 mmol) of1-formyl-1-methylcyclohexane. 25.9 g of 4-methylcyclohexylglycinehydrochloride were obtained starting from 16.8 g (150 mmol) of4-methylcyclohexanone. 18 g of trans-4-methyl-1-cyclohexylalaninehydrochloride were obtained starting from 15 g oftrans-1-formyl-4-methylcyclohexane. 10 g of3,3-dimethyl-1-cyclohexylalanine hydrochloride were obtained startingfrom 9 g of 3,3-dimethyl-1-formylcyclohexane.

The aldehyde 1-formyl-3,3-dimethylcyclohexane required for the synthesiswas prepared by a method based on that of Moskal and Lensen (Rec. Trav.Chim. Pays-Bas 106 (1987) 137-141).

A solution of n-butyllithium in n-hexane (72 ml, 115 mmol) was addeddropwise over the course of 10 min to a stirred solution of diethylisocyanomethylphosphonate (17 ml, 105 mmol) in 280 ml of anhydrousdiethy [sic] ether at −60° C. The resulting suspension was then stirredat −60° C. for 15 min and, over the course of 10 min, a solution of3,3-dimethylcyclohexanone (13 g, 105 mmol) in 100 ml of anhydrousdiethyl ether was added, keeping the temperature below −45° C. Thereaction mixture was allowed to reach 0° C. and, after stirring at thistemperature for 90 min, 150-200 ml of 38% strength aqueous hydrochloricacid were cautiously added. The mixture was vigorously stirred at roomtemperature for 15 h to complete the hydrolysis. The organic phase wasseparated off and washed with 200 ml each of water, saturated sodiumbicarbonate solution and saturated sodium chloride solution. It wasdried over magnesium sulfate, filtered and concentrated in a rotaryevaporator in order to remove the solvent. The resulting residue wasemployed without further purification as starting material forsynthesizing the amino acid.

Cyclopentylglycine was prepared by hydrolysingN-acetyl-(D,L)-cyclopentylglycine with 6N hydrochloric acid, the formerhaving been prepared as described in the literature by J. T. Hill and F.W. Dunn, J. Org. chem. 30(1965), 1321.

Boc-(D)-α-Methylcyclohexylalanine

3.4 g (12.2 mmol) of Boc-(D)-α-methyl-Phe-OH were hydrogenated in 100 mlof MeOH in the presence of 250 mg of 5% Rh on Al₂O₃ under 10 bar withhydrogen at 50° C. for 24 h. Filtration and stripping off the solventresulted in 2.8 g of Boc-(D)-α-methyl-Cha-OH.

¹H NMR (DMSO-d₆, δ in ppm): 12 (very broad signal, COOH); 1.7-0.8 (25H;1.35 (s, Boc), 1.30 (s, Me)).

Boc-(3-Ph)-Pro-OH was synthesized by a method similar to that of J. Y.L. Chung et al. (J. Y. L. Chung et al., J.Org.Chem. 55 (1990) 270).

Preparation of Boc-1-Tetralinylglycine

Boc-1-Tetralinylglycine was prepared starting from1,2-dihydronaphthalene. 1,2-Dihydronaphthalene was initially convertedinto 1-tetralyl bromide with HBr (similar to J. Med. Chem. 37 (1994)1586). The bromide was subsequently reacted with diethylacetamidomalonate and, after hydrolytic cleavage, the resulting α-aminoacid was converted into the Boc-protected form under standardconditions. Another possible preparation is described by E. Reimann andD. Voss (E. Reimann, D. Voss, Arch. Pharm. 310 (1977) 102).

Preparation of Boc-(D,L)Dch-OH

Boc-(D,L)-Dpa-OH (1 mmol) was hydrogenated in 12 ml of MeOH togetherwith catalytic amounts of 5% Rh/Al₂O₃ under 5 bar. Filtration andremoval of the solvent under reduced pressure resulted in the product inquantitative yield. Preparation of H-D,L-Chea-OH

4.0 g of cycloheptylmethyl methanesulfonate (19.39 mmol), prepared fromcycloheptylmethanol and methanesulfonyl chloride, were refluxed togetherwith 4.9 g of benzophenone imine glycine ethyl ester (18.47 mmol), 8.9 gof dry, finely powdered potassium carbonate (64.65 mmol) and 1 g oftetrabutylammonium bromide (3 mmol) in 50 ml of dry acetonitrile underan inert gas atmosphere for 10 h. The potassium carbonate was thenfiltered off, the filtrate was evaporated to dryness, and the crudeproduct was hydrolyzed directly with 20 ml of 2N hydrochloric acid in 40ml of ethanol, stirring at RT for 1.5 h. The reaction solution wasdiluted and then benzophenone was extracted with ethyl acetate in theacidic range, and subsequently H-D,L-Chea-OEt was extracted with DCM inthe alkaline range (pH=9), and the solution was dried over magnesiumsulfate and concentrated in a rotary evaporator. Yield 3.7 g{circumflexover (=)}95% of theory.

Boc-(D,L)-(3,4,5-(MeO)₃)Phe-OH was prepared by alkylation ofbenzophenone imine glycine ethyl ester with trimethoxybenzyl chloride,subsequent introduction of the Boc protective group and esterhydrolysis.

H-(D,L)-P,P-Me₂Cha-OH was prepared by the method of U. Schöllkopf, R.Meyer, L. Ann. Chem. (1977) 1174-82.

Said amino acids were converted with di-tert-butyl dicarbonate inwater/dioxane by conventional methods into the Boc-protected form ineach case and subsequently recrystallized from ethyl acetate/hexanemixtures or purified by column chromatography on silica gel (mobilephases: ethyl acetate/petroleum ether mixtures).

The Boc-protected amino acids were employed as B building blocks asshown in Scheme I.

Said amino acids as B building blocks were also in some cases convertedinto the corresponding benzyl esters and linked to the appropriatelyprotected A building blocks. In the case of compounds with an N—Hfunctionality which was still free, this was subsequently protected witha Boc group, the benzyl ester group was removed by hydrogenation, andthe building block A—B—OH was purified by crystallization, saltprecipitation or column chromatography. This route is described by wayof example for tBuOOC—CH₂—(Boc)(D)Cha-OH below.

Synthesis of (D)-Cyclohexylalanine Benzyl Ester

A suspension of 100 g (481 mmol) of (D)-cyclohexylalanine hydrochloride,104 g (962 mmol) of benzyl alcohol and 109.7 g (577 mmol) ofp-toluenesulfonic acid monohydrate in 2200 ml of toluene was slowlyheated to reflux with a water separator. Evolution of hydrogen chlorideand dissolving of the suspension to give a clear solution were observedin the temperature range 80-90° C. When no further water separated out(about 4 h), 500 ml of toluene were distilled out, the reaction mixturewas allowed to cool overnight, and the resulting residue was filteredoff and washed twice with 1000 ml of hexane each time. The resultingresidue (195 g) was then suspended in 2000 ml of dichloromethane and,after addition of 1000 ml of water, adjusted to pH 9-9.5 by gradualaddition of 50% strength sodium hydroxide solution while stirring. Theorganic phase was separated off, washed twice with 500 ml of water eachtime, dried over sodium sulfate and filtered to remove desiccant, andconcentration of the filtrate resulted in 115 g (94%) of the titleproduct as pale oil.

N-(tert-Butyloxycarbonylmethylene)-(D)-cyclohexylalanine Benzyl Ester

115 g (440 mmol) of (D)-cyclohexylalanine benzyl ester were dissolved in2000 ml of acetonitrile and, at room temperature, 607.5 g (4.40 mol) ofpotassium carbonate and 94.3 g (484 mmol) of tert-butyl bromoacetatewere added, and the mixture was stirred at this temperature for 3 days.Carbonate was filtered off, washed with acetonitrile, the mother liquorwas concentrated (30° C., 20 mbar), the residue was taken up in 1000 mlof methyl tert-butyl ether, and the organic phase was extracted with 5%strength citric acid and saturated sodium bicarbonate solution. Theorganic phase was dried over sodium sulfate, filtered to removedesiccant and concentrated, and the resulting oil (168 g) was employeddirectly in the next reaction.

N-Boc-N-(tert-Butyloxycarbonylmethylene)-(D)-cyclohexylalanine BenzylEster

The oil (168 g, 447 mmol) obtained in the previous synthesis wasdissolved in 1400 ml of acetonitrile and, after addition of 618 g (4.47mmol) of potassium carbonate powder and 107.3 g (492 mmol) ofdi-tert-butyl dicarbonate, stirred at room temperature for 6 days. Thepotassium carbonate was filtered off with suction, washed with about1000 ml of acetonitrile, and the filtrate was concentrated. 230 g of therequired product were obtained.

N-Boc-N-(tert-Butyloxycarbonylmethylene)-(D)-cyclohexylalanineCyclohexylammonium Salt

115 g of N-Boc-N-(tert-butyloxycarbonylmethylene)-(D)-cyclohexylalaine[sic] benzyl ester were dissolved in 1000 ml of pure ethanol andhydrogenated in the presence of 9 g of 10% Pd on active carbon withhydrogen under atmospheric pressure at 25-30° C. for 2 h. Filtration andremoval of the solvent in a rotary evaporator resulted in 100 g (260mmol) of a yellow oil which was taken up in 1600 ml of acetone andheated to reflux. The heating bath was removed, and a solution of 27 g(273 mmol) of cyclohexylamine in acetone was quickly added through adropping funnel. The required salt crystallized out on cooling thereaction mixture to room temperature. The solid was filtered off, washedwith 200 ml of acetone and, for final purification, recrystallized oncemore from acetone. Drying of the residue in a vacuum oven at about 30°C. resulted in 70.2 g of the required salt as white powder.

N-Boc-N-(tert-Butyloxycarbonylmethylene)-(D)-cyclohexylglycinecyclohexylammonium salt was prepared analogously from cyclohexylglycineas precursor. TheN-Boc-N-(tert-butyloxycarbonylmethylene)-(D)-cycloheptylglycine andN-Boc-N-(tert-butyloxycarbonylmethylene)-(D)-cyclopentylglycinederivatives were prepared from the corresponding cycloheptyl- andcyclopentylglycine compounds.

N-Boc-N-(tert-Butyloxycarbonylethylene)-(D)-cyclohexylalanineCyclohexylammonium Salt

a) tert-Butyl 3-Bromopropionate

In a countercurrent of nitrogen, 16.64 g (109 mmol) of bromopropionicacid, 150 ml of condensed 2-methylpropene and 2 ml of concentratedsulfuric acid were added at −30° C. into an autoclavable glass vessel,the autoclave was sealed tightly and the mixture was stirred for 72hours at room temperature. For working up, the reaction vessel was againcooled to −30° C. and the reaction solution was poured carefully into200 ml of an ice-cold saturated sodium hydrogen carbonate solution.Excess 2-methylpropene was evaporated with stirring, the residue wasextracted three times with in each case 50 ml of dichloromethane, andthe combined organic phases were dried over sodium sulfate, filtered toremove desiccant and concentrated under a water pump vacuum. The oilyresidue was purified by column chromatography (mobile phase n-hexane,later n-hexan/diethyl ether 9:1). This resulted in 18.9 g of the titlecompound.

b) N-(tert-Butyloxycarbonylethylene)-(D)-cyclohexylalanine Benzyl Ester

49.4 g (189 mmol) of (D)-cyclohexylalanine benzyl ester were dissolvedin 250 ml of acetonitrile, the solution was treated with 31.6 g (151mmol) of tert-butyl bromopropionate at room temperature and the mixturewas refluxed for 5 days. The mixture was filtered to remove theprecipitate formed and this was washed repeatedly with acetonitrile, thefiltrate was concentrated under a water pump vacuum, the residue wastaken up in 350 ml of dichloromethane, and the organic phase wasextracted with 5% strength citric acid and saturated sodium hydrogencarbonate solution. The organic phase was dried over sodium sulfate,filtered to remove desiccant and concentrated. The oily residue waspurified by column chromatography (mobile phase dichloromethane, laterdichloromethane/methanol 95:5). This resulted in a slightly impure oilwhich was employed directly in the next reaction.

c) N-Boc-N-(tert-Butyloxycarbonylethylene)-(D)-cyclohexylalanine BenzylEster

The oil obtained in the synthesis above (30 g, max. 70 mmol) wasdissolved in 150 ml of acetonitrile, and the solution was treated with28 ml (160 mmol) of diisopropylethylamine and 19.2 g (88 mmol) ofdi-tert-butyl dicarbonate and stirred for 3 days at room temperature.The reaction mixture was concentrated under a water pump vacuum in arotary evaporator, the residue was taken up in n-hexane, the mixture waswashed 5 times using in each case 3 ml of a 5% strength citric acidsolution, the combined organic phases were dried over sodium sulfate,filtered to remove desiccant and concentrated, and the residue wassubjected to separation by column chromatography (mobile phasehexane/ethyl acetate 95:5). The results were 32.66 g (64 mmol) of therequired product.

d) N-Boc-N-(tert-Butyloxycarbonylethylene)-(D)-cyclohexylalanineCyclohexylammonium Salt 32.66 g (64 mmol) ofN-Boc-N-(tert-butyloxycarbonylethylene)-(D)-cyclohexylalanine benzylester were dissolved in 325 ml of pure ethanol and hydrogenated withhydrogen for 14 hours under atmospheric pressure at 25-30° C. in thepresence of 3 g of 10% pure Pd on active charcoal. Filtration of thesolution through Celite®, washing of the latter with ethanol and removalof the solvent in a rotary evaporator resulted in 26.7 g of a yellow oilwhich was taken up in acetone and heated to reflux. The heating bath wasremoved, and a solution of 7 g (70 mmol) of cyclohexylamine in acetonewas quickly added through a dropping funnel. The required saltcrystallized out on cooling the reaction mixture to room temperature.The solid was filtered off, washed with 25 ml of acetone and, for finalpurification, recrystallized once more from acetone. Drying of theresidue in a vacuum oven at 30° C. resulted in 26.6 g (54 mmol) of therequired salt as white powder.

N-Boc-N-(tert-butyloxycarbonylmethylene)-(D)-cyclohexylalanyl-3,4-dehydroproline:

a) N-Boc-Pyr-OH (5 g, 23.45 mmol) was dissolved in MeOH (50 ml), and HClin dioxane (4N, 30 ml) was added. The mixture was subsequently heatedunder reflux for 12 hours. Removal of the solvent in a rotary evaporatorresulted in H-Pyr-OMe-hydrochloride as product. Yield: 3.84 g (100%).

b) N-(t-Bu02C—CH₂)-N-Boc-(D)-Cha-OH (8 g, 20.75 mmol) was dissolved indichloromethane (75 ml), and ethyldiisopropylamine (15.5 ml, 89.24 mmol)were added at −10° C. After the mixture had been stirred for 5 minutesat this temperature, a solution of H-Pyr-OMe hydrochloride (3.4 g, 20.75mmol) in dichloromethane (25 ml) was added dropwise. A solution ofpropanephosphonic anhydride in ethyl acetate (50% strength, 20 ml, 26.96mmol) was subsequently added dropwise and stirred for 2 h at −10 to 0°C. The batch was diluted with dichloromethane and washed with saturatedsodium hydrogen carbonate solution (2×80 ml), 5% strength citric acidsolution (2×15 ml) and saturated sodium chloride solution (1×20 ml). Theorganic phase was dried over sodium sulfate and the solvent was removedin a rotary evaporator. The crude product was purified by means of flashchromatography (silica gel, dichloromethane/methanol 95/5). Yield: 6.2 g(60%).

c) N-(t-BuO₂C—CH₂)-N-Boc-(D)-Cha-Pyr-OMe (5.5 g, 11.12 mmol) wasdissolved in dioxane (40 ml), aqueous sodium hydroxide solution (IN,22.2 ml, 22.24 mmol) was added, and the mixture was stirred for 2 hoursat room temperature. The dioxane was removed in a rotary evaporator, andthe aqueous phase was washed with ethyl acetate and acidified to pH 1-2with potassium hydrogen sulfate solution (20% strength). The aqueousphase was extracted with dichloromethane and the combined organic phaseswere dried over sodium sulfate. Yield: 5 g (94%), colorless foam.Recrystallization of n-hexane saturated with water resulted in thecorresponding carboxylic acid as colorless crystals (m.p.=158-160° C.).

N-Boc-N-(tert-butyloxycarbonylmethylene)-(D)-cyclohexylglycyl-3,4-dehydroproline:

This compound was synthesized analogously fromN-Boc-N-(tert-butyloxycarbonylmethylene)-(D)-cyclohexylglycine and3,4-dehydroproline methyl ester.

N-Boc-N-(tert-butyloxycarbonylmethylene)-(D)-cyclohexylalanylproline:

a) N-(t-BuO₂C—CH₂)-N-Boc-(D)-Cha-OH (20 g, 51.88 mmol) was dissolved indry methylene chloride (100 ml). After cooling to −5° C.,N-ethyldiisopropylamine (90 ml, 518.88 mmol) was added dropwise andstirring was continued for 5 minutes.

H-Pro-OBn×HCl (12.54 g, 51.88 mmol) was subsequently added at −5° C.and, after the mixture had been stirred for 5 minutes, 50% strengthpropanephosphonic anhydride solution in ethyl acetate (45.1 ml, 62.26mmol), diluted with methylene chloride (45 ml), was added dropwise inthe course of 30 minutes. After the mixture had been stirred for 1 hourat 0-5° C., it was slowly brought to RT and stirred for 12 hours at RT.The batch was diluted with methylene chloride and washed in successionwith saturated sodium hydrogen carbonate solution, 5% strength citricacid solution and saturated sodium chloride solution. After drying oversodium sulfate, the solvent was distilled off in vacuo. Yield: 28.9 g(pale yellow oil, 97%).

b) The product obtained as described in a) (28.5 g, 49.76 mmol) wasdissolved in methanol (650 ml), 10% pure Pd on charcoal (1.8 g) wasadded, and the mixture was hydrogenated at RT and under 1 atmosphere ofhydrogen. The catalyst was subsequently removed by filtration throughCelite® and the filtrate was concentrated in vacuo. Yield: 22.2 g(colorless foam, 92%).

N-Boc-N-(tert-butyloxycarbonylmethylene)-(D)-cyclohexylglycylproline:

This compound was prepared analogously fromN-Boc-N-(tert-butyloxycarbonylmethylene)-(D)-cyclohexylglycine andproline methyl ester.

D building blocks:

The compounds employed as D building blocks, (L)-proline, (L)-pipecolicacid and (L)-azetidinecarboxylic acid, are commercially available,either as free amino acids, as Boc- protected compounds or as thecorresponding methyl esters. If (L)-3,4-dehydroproline or(D,L)-4,5-dehydropipecolic acid, or a corresponding, protectedderivative, were employed as D building blocks, the compounds preparedwere generally hydrogenated in the last step to give the correspondingproline derivatives. (L)-3,4-Dehydroproline (H-Pyr-OH) is commerciallyavailable, (D,L) 4,5-dehydropipecolic acid (H-(D,L)-Dep-OH) can beprepared by the methods of A. Burgstahler, C. E. Aiman J. Org. Chem. 25(1960), 489 or C. Herdeis, W. Engel Arch. pharm 326 (1993), 297.

The E building blocks were synthesized as follows:

5-Aminomethyl-2-cyanothiophene:

This building block was synthesized as described in WO 95/23609.

4-Aminomethyl-2-cyanothiophene

a) 2-Bromo-4-formylthiophene

36 g (320 mmol) of 3-formylthiophene were dissolved in 600 ml ofmethylene chloride, the solution was cooled to 5° C., 100 g (750 mmol)of aluminum trichloride were added, a little at a time, and the reactionmixture was subsequently refluxed. A solution of 59 g (19 ml, 360 mmol)of bromine in 40 ml of methylene chloride was added dropwise over thecourse of 45 minutes and the mixture was allowed to after-react underreflux for 4 hours. After cooling, the reaction solution was poured onto600 g of ice-water and extracted with methylene chloride, and theorganic phase was washed with saturated sodium hydrogen carbonatesolution, dried over magnesium sulfate and concentrated in a rotaryevaporator in vacuo. 64.5 g of crude product resulted, which waspurified by means of column chromatography (silica gel, methylenechloride/petroleum ether). A total of 56.5 g of slightly impure productwere obtained.

b) 2-Cyano-4-formylthiophene

7.6 g (85 mmol) of copper(I) cyanide were added to a solution of 13.53 g(70.82 mmol) of 2-bromo-4-formylthiophene in 25 ml of DMF and thereaction mixture was refluxed for 3.5 hours, during which process thesuspension, which originally was pale green in color, turned into ablack solution. After addition of water, the reaction mixture wasextracted repeatedly with ethyl acetate, and the organic phases werecombined, washed with saturated sodium chloride solution, dried oversodium sulfate and concentrated under slightly reduced pressure.Treatment of the residue (7 g) with ether resulted in 1.6 g of pureproduct. The mother liquor together with the crude products from otherbatches was purified by chromatography. (Silica. gel, methylenechloride/petroleum ether 1:1). In total, 56.5 g of2-bromo-4-formylthiophene were reacted to give2-cyano-4-formylthiophene, resulting in 12.6 g of pure product (yield31%).

c) 2-Cyano-4-hydroxymethylthiophene

3.47 g (91.8 mmol) of sodium borohydride were added, a little at a time,to a suspension of 12.6 g (91.8 mmol) of 2-cyano-4-formylthiophene in200 ml of ethanol and stirred at room temperature for 2 hours, duringwhich process the reaction mixture slowly formed a clear solution. Afterconcentration in vacuo, the residue was taken up in ethyl acetate,washed in succession with saturated sodium chloride solution, 5%strength citric acid and saturated sodium chloride solution, and theorganic phase was dried using sodium sulfate and concentrated in vacuo.11.7 g of almost pure product resulted (yield 91.5%).

d) 4-Bromomethyl-2-cyanothiophene

11.7 g (84.07 mmol) of 2-cyano-4-hydroxymethylthiophene together with24.1 g (91.87 mmol) of triphenylphosphine were dissolved in 100 mlof-THF at room temperature, and 30.47 g (91.87 mmol) oftetrabromomethane were added, a little at a time, with cooling (icebath). After stirring for 3 hours at room temperature, the mixture wasconcentrated in vacuo and purified by chromatography over silica gel(methylene chloride/petroleum ether). 18.8 g of crystalline pale yellowproduct which still contained petroleum ether resulted.

e) 4-N,N-bis(tert-Butoxycarbonyl)aminomethyl-2-cyanothiophene

18.81 g of 4-bromomethyl-2-cyanothiophene (crude product, maximum 84.07mmol) were dissolved in 160 ml of THF, the solution was cooled to 5° C.,and 3.07 g (102.4 mmol) of 80% sodium hydride suspension were added, alittle at a time. 22.25 g (102.4 mmol) of di-tert-butyliminodicarboxylate, dissolved in 160 ml of THF, were subsequently addeddropwise at 5° C. and the mixture was then stirred overnight at roomtemperature. Since TLC revealed that the reaction was incomplete, thebatch was heated for 4.5 hours at 30-35° C.

After cooling to 0-5° C., 33 ml of saturated ammonium chloride solutionwere slowly added dropwise, THF was distilled off in vacuo, the residuewas extracted repeatedly with ethyl acetate, and the ethyl acetatephases were washed with saturated sodium chloride solution, dried oversodium sulfate and evaporated in a rotary evaporator. The red viscousresidue (34.61 g) was employed in the subsequent reaction as crudeproduct.

f) 4-Aminomethyl-2-cyanothiophene hydrochloride

34.61 g of 4-N,N-bis(tert-butoxycarbonyl)aminomethyl-2-cyano-thiophene(crude product, maximum 84.07 mmol) were dissolved in 600 ml of ethylacetate, and the solution was cooled to 0-5° C., saturated with HCl gasand warmed to room temperature. After 3 hours, the resulting suspensionwas evaporated in a rotary evaporator, the product was codistilledrepeatedly with methylene chloride, and the residue was extracted bystirring with ether and dried in vacuo. 13.85 g of product resulted as apale powder. Yield over two steps: 94.3%.

2-Aminomethyl-4-cyanothiophene

a) 4-Cyanothiophene-2-carbaldehyde

49.3 g (258.05 mmol) of 4-bromothiophene-2-carbaldehyde and 27.8 g(310.41 mmol) of copper(I) cyanide were suspended in 130 ml of absoluteDMF and the suspension was refluxed for 8 hours. The solvent wasevaporated in vacuo in a rotary evaporator at 400C, the residue wassuspended in ethyl acetate and the suspension was transferred into aSoxleth apparatus. The residue was extracted overnight, the yellowsolution was dried over sodium sulfate and evaporated in vacuo in arotary evaporator, and the resulting yellow solid was recrystallizedfrome ether. 25.3 g of product resulted (80% of theory).

b) 4-Cyanothiophene-2-carbaldehyde oxime

11.6 g (84.6 mmol) of 4-cyanothiophene-2-carbaldehyde were dissolved in140 ml of methanol and 12.3 g (116.1 mmol) of sodium carbonate wereadded. 6.5 [lacuna] (93.5 mmol) of hydroxylamine hydrochloride weresubsequently added at 15° C. with cooling, a little at a time, and themixture was stirred for 2 hours at 10° C. After 80 ml of water had beenadded, the reaction mixture was extracted five times using in each case50 ml of diethyl ether, the organic phase was dried over sodium sulfateand the solvent was removed in vacuo. 12.5 g of the required productresulted as a yellow crystal powder (96% of theory).

c) 2-Aminomethyl-4-cyanothiophene Hydrochloride

11.22 g (171.64 mmol) of fine zinc dust were carefully added, in severalsmall portions, to a solution of 4.65 g (30.60 mmol) of4-cyanothiophene-2-carbaldehyde oxime in 50 ml of trifluoroacetic acid,cooled to 0-5° C., in such a way that the temperature did not climbabove 15° C. After stirring for 3 hours at room temperature, excess zincwas decanted off, most of the trifluoroacetic acid was removed in vacuo(oil pump), the remaining oil was cooled to 0° C., and a mixture of 150ml of 3N aqueous sodium hydroxide solution and 21 of methylene chloride,pre-cooled to 0° C., was added, a little at a time. After insolubleswere removed by filtration, the organic phase was separated off, theaqueous phase was extracted eight times using 20 ml of methylenechloride, the collected organic phases were dried over sodium sulfate,and 20 ml of 6M methanolic hydrochloric acid were subsequently added,with ice-cooling. During this process, the product precipitated in theform of the hydrochloride as a white solid, the suspension being cooledovernight to 4° C. to bring crystallization to completion. 2.2 g ofproduct resulted as colorless needles (50% of theory).

5-Aminomethyl-3,4-dimethylthiophene-2-carboxamide Hydrochloride

19 g (105.42 mmol) of 5-cyano-3,4-dimethylthiophene-2-carboxamide weresuspended in 760 ml of methanol and 110 ml of 2N hydrochloric acidsolution, 9.5 g of Pd on charcoal (10%) were added, and the mixture washydrogenated at room temperature. After 4.71 of hydrogen had been takenup (4 h), methanol was distilled out in vacuo, and the aqueous phase wasextracted three times with ethyl acetate and subsequently freeze-dried.16.3 g of the required product resulted as a white solid (70.4% oftheory).

5-Aminomethylisoxazole-3-carboxamide

a) Ethyl 5-Chloromethylisoxazole-3-carboxylate

21.2 g (210 mmol) of triethylamine were added dropwise with stirring toa mixture, cooled to 10-15° C., of 30 g (198 mmol) of ethyl2-chloro-2-hydroxyiminoacetate and 150 ml of propargyl chloride,stirring was continued for 1 hour at room temperature, water wassubsequently added, the mixture was extracted with ether, and theorganic phase was dried over magnesium sulfate and evaporated in vacuoin a rotary evaporator. The residue was distilled in vacuo at 0.5 torr,the product distilling over at 116-122° C.

b) 5-Chloromethylisoxazole-3-carboxylic Acid

14 g (250 mmol) of potassium hydroxide were added to 47.3 g (250 mmol)of ethyl 5-chloromethylisoxazole-3-carboxylate in 150 ml of ethanol, andthe reaction mixture was stirred for 6 hours at 60-70° C. After cooling,the mixture was concentrated in vacuo, the residue was taken up in waterand extracted with ether, the aqueous phase was acidified withhydrochloric acid and subsequently extracted repeatedly with ether, andthe ether phase was dried over sodium sulfate and concentrated in vacuo(oil pump, 50° C.). 31 g of the required product resulted (77% oftheory)

c) 5-Chloromethylisoxazole-3-carboxylic Acid Chloride

120 g (743 mmol) of 5-chloromethylisoxazole-3-carboxylic acid togetherwith 500 ml of thionyl chloride and 2 drops of pyridine were refluxedfor 10 hours, subsequently concentrated in vacuo and then distilled at20 torr. The product distilled at 125-1330C. 78 g resulted (58% oftheory)

d) 5-Chloromethylisoxazole-3-carboxamide

Ammonia was passed for 1 hour at 10-15° C. into a solution of 10 g(55.56 mmol) of 5-chloromethylisoxazole-3-carboxylic acid chloride in100 ml of methylene chloride and stirring was subsequently continued atroom temperature for 1 hour. After the solution had cooled to 0° C., theprecipitate was filtered off with suction and washed with a little coldmethylene chloride, and the residue was extracted twice by stirring withwater to remove the ammonium salts. Drying in vacuo resulted in 6.58 gof pure product as a pale powder (74% of theory)

e) 5-Aminomethylisoxazole-3-carboxamide Hydrochloride

2.44 g (15.2 mmol) of 5-chloromethylisoxazole-3-carboxamide were addedto a mixture of 100 ml of concentrated ammonia solution and 72 ml ofmethanol, and the reaction solution was warmed to 40° C. and constantlysaturated with ammonia gas during this process. After 6 hours, theprecursor was reacted. The methanol was removed in vacuo, and theaqueous phase was extracted twice using methylene chloride andsubsequently evaporated to dryness in vacuo under mild conditions in arotary evaporator. The white solid residue was employed in the couplingreactions in the form of the crude product.

2-Aminomethyloxazole-4-thiocarboxamide and2-aminomethylthiazole-4-thiocarboxamide were prepared by the method ofG. Videnov, D. Kaier, C. Kempter and G. Jung Angew. Chemie (1996) 108,1604, where the N-Boc-protected compounds described therein weredeprotected using etheric hydrochloric acid in methylene chloride.

4-Aminomethylthiazole-2-thiocarboxamide

a) Monothiooxalic Diamide

Monothiooxalic diamide was prepared starting from ethyl thiooxamidate bythe method of W. Walter, K.-D. Bode Liebigs Ann. Chem. 660 (1962),74-84.

b) 2-Carbamoyl-4-chloromethylthiazole

10 g (96 mmol) of ethyl thiooxamidate were introduced into 170 ml ofn-butanol, 26 g (204 mmol) of 1,3-dichloroacetone were added, and themixture was heated for 90 minutes at 112° C. under nitrogen. Thereaction mixture was then concentrated in vacuo and the residue wasextracted by stirring with n hexane [sic] (120 ml). 10 g of pure productresulted.

c) 4-Boc-Aminomethyl-2-carbamoylthiazole

10 g (56.6 mmol) of 2-carbamoyl-4-chloromethylthiazole were introducedinto an ammonia-saturated solution of 350 ml of methanol and 80 ml of25% strength aqueous ammonia solution. The reaction mixture was warmedfor 6 hours at 40-42° C. while continuously saturating with ammonia,then concentrated in vacuo and codistilled with methanol, and theresidue was subsequently extracted by stirring first with ether and thenwith acetone. 7.6 g of crude product which still contained a smallamount of ammonium chloride were isolated. To remove this secondaryproduct, the crude product was reacted with (Boc)₂O in aqueous dioxanesolution, and the protected compound was purified by means of columnchromatography. This resulted in 4.95 g of pure product.

d) 4-Boc-Aminomethyl-2-cyanothiazole

4.95 g (19.24 mmol) of 4-Boc-aminomethyl-2-carbamoylthiazole wereintroduced into 90 ml of methylene chloride and 16.7 ml (97.44 mmol) ofdiisopropylethylamine, the mixture was cooled to 0° C., a solution of6.35 ml of trifluoroacetic anhydride in 10 ml of methylene chloride wasadded dropwise at 0 to 5° C., and the mixture was subsequently warmed toroom temperature (TLC check). Then, 25 ml of water were added, themixture was stirred for 30 minutes at room temperature and brought to pH2.5 with 10% strength citric acid solution, and the organic phase waswashed repeatedly, dried using magnesium sulfate and concentrated invacuo. 5.4 g of viscous, pale brown crude product resulted, which wereemployed in the next step without further purification.

e) 4-Boc-Aminomethyl-2-thiocarbambylthiazole

The crude product resulting from d) (max 19.24 mmol) was dissolved in 65ml of pyridine and 5 ml of triethylamine, saturated with hydrogensulfide and left to stand at room temperature over the weekend. Thereaction mixture was then evaporated in vacuo in a rotary evaporator,the residue was taken up in a mixture of ether and ethyl acetate, andthe mixture was washed with 10% strength citric acid solution and water,dried over magnesium sulfate and evaporated in vacuo in a rotaryevaporator. 6.0 g resulted as a pale yellow solid foam.

f) 4-Aminomethyl-2-thiocarbamoylthiazole Hydrochloride

The product resulting from the above experiment was taken up in 100 mlof methylene chloride, 30 ml of approx. 5-molar etheric hydrochloricacid solution were added, and the mixture was stirred overnight at roomtemperature. The reaction mixture was then evaporated to dryness invacuo in a rotary evaporator, codistilled repeatedly with ether andsubsequently extracted by stirring with methylene chloride. 4.15 g ofthe required product resulted as a pale yellow amorphous substance.

4-Amidino-2-(N-Boc-aminomethyl)-5-methylthiazole×HOAc

a) α-Acetylglycine Methyl Ester Hydrochloride

Potassium tert-butylate (17.8 g, 157.9 mmol) was introduced into THF(120 ml), and a solution of N-diphenylmethylideneglycine methyl ester(40 g, 157.9 mmol) in THF (60 ml) was added at −70° C. After theyellowish solution had been stirred for 30 minutes at this temperature,it was added dropwise at −70° C. to a solution of acetyl chloride (12.4g, 157.9 mmol) in THF (70 ml). After the mixture had been stirred atthis temperature for 1.75 hours, 3N HCl (160 ml) was added, and theyellowish suspension was stirred for a further 10 minutes at roomtemperature. The THF was removed at room temperature on a rotaryevaporator, and the remaining aqueous phase was washed 3× with diethylether. The aqueous phase was freeze-dried and the residue was extractedby stirring with methanol. The methanolic solution of the product wasconcentrated on a rotary evaporator at 35° C. Yield: 26.4 g (157.9 mmol,quant., yellowish solid).

b) BOC-Gly-(α-Acetyl-Gly)-OMe [sic]

BOC-Gly-OH [sic] (24.05 g, 137.27 mmol) were [sic] introduced into THF(400 ml), and triethylamine (13.87 g, 137.19 mmol) was added. Thecolorless solution was cooled to −20° C., and a solution of isobutylchloroformate (18.75 g, 137.28 mmol) in THF (20 ml) was added dropwiseat this temperature. The colorless suspension was stirred for a further30 minutes at −20° C., and a-acetylglycine methyl ester hydrochloride(23.0 g, 137.3 mmol) was then added portionwise. After the mixture hadbeen stirred for 30 minutes at −20° C., a solution of triethylamine(13.87 g, 137.19 mmol) in THF (20 ml) was added dropwise in the courseof 45 minutes. After the mixture had been stirred for 4 hours at −20°C., stirring was continued for another 12 hours at RT. The residue wasfiltered off with suction and-washed with THF, and the combined THFphases were concentrated on a rotary evaporator.

Yield: 44.1 g (pale brown oil). ¹H NMR (270 MHz, CDCl₃) δ=1.45 (s, 9H),2.40 (s, 3H), 3.85 (s, 3H), 3.90 (d, J=6.5 Hz, 2H), 5.25 (d, J=6.5 Hz,1H), 7.30 (sbr, 1H).

c) Methyl 2-(N-Boc-Aminomethyl)-5-methylthiazole-4-carboxylate

BOC-Gly-(a-acetyl-Gly)-OMe [sic] (39.8 g, 138.2 mmol) was introducedinto THF (400 ml), and Lawesson's reagent (96.6 g, 238.8 mmol) was addedportionwise at room temperature. The yellowish solution was thenrefluxed for 1.5 hours. The THF was removed on a rotary evaporator. Theresidue (reddish-brown oil) was extracted by stirring with diethyl ether(600 ml). The ether phase was decanted off from the undissolved brownishoil .and washed in succession with 5% strength citric acid (2×),saturated NaHCO₃ solution (9×) and water (2×). After drying (MgSO₄) thesolvent was removed on a rotary evaporator. Yield: 22.0 g (77 mmol, 56%,brownish solid).

¹H NMR (270 MHz, CDCl₃) δ=1.50 (s, 9H), 2.75 (s, 3H), 3.95 (s, 3H), 4.55(d, J=6.5 Hz, 2H), 5.45 (t, J=6.5 Hz, 1H). (Main rotamer relative to theBoc group).

d) 2-(N-Boc-Aminomethyl)-5-methylthiazole-4-carboxylic Acid

Methyl 2-(N-Boc-aminomethyl)-5-methylthiazole-4-carboxylate (22.0 g, 77mmol) was dissolved in ethanol (100 ml), and a solution of LiOH (2.2 g,92 mmol) in water (50 ml) was added. After the mixture had been stirredfor 30 minutes at room temperature, the ethanol was removed on a rotaryevaporator and the solution which remained was diluted with water (70ml). The aqueous phase was washed with ethyl acetate (3×) and brought topH 2 with 20% strength NaHSO₄ solution, during which process a palebrown oil separated out. The aqueous phase was extracted withdichloromethane and the combined organic extracts were dried (MgSO₄) andconcentrated in vacuo. The pale brown residue was extracted by stirringin diisopropyl ether. The colorless precipitate which remained wasfiltered off with suction and washed with diisopropyl ether. Yield: 6.9g (25.4 mmol, 33%, colorless solid).

¹H NMR (270 MHz, DMSO-d₆) δ=1.40 (s, 9H), 2.65 (s, 3H), 4.30 (d, J=6.5Hz, 2H), 7.80 (t, J=6.5 Hz, 1H).

e) 2-(N-Boc-Aminomethyl)-5-methylthiazole-4-carboxamide

2-(N-Boc-Aminomethyl)-5-methylthiazole-4-carboxylic acid (6.8 g, 25mmol) was dissolved in THF (100 ml), and triethylamine (2.53 g, 25 mmol)was added. After the mixture had been cooled to −20° C., a solution ofisobutyl chloroformate (3.41 g, 25 mmol) in THF (10 ml) was addeddropwise. After the mixture had been stirred for 30 minutes at −20° C.,ammonia gas was passed into the pale brown suspension for 45 minutes.The mixture was then warmed to room temperature. The residue wasfiltered off with suction and extracted with THF, and the filtrates wereconcentrated.

Yield: 6.9 g (25 mmol, quant.). ¹H NMR (270 MHz, DMSO-d₆) δ=1.40 (s,9H), 2.65 (s, 3H), 4.30 (m, 2H), 7.40 (sbr, 1H), 7.50 (sbr, 1H), 7.80(t, J=6.5 Hz, 1H).

f) 4-Cyano-2-(N-Boc-aminomethyl)-5-methylthiazole

2-(N-Boc-Aminomethyl)-5-methylthiazole-4-carboxamide (6.8 g, 25 mmol)was introduced into dichloromethane (120 ml). After the mixture hadcooled to 0° C. diisopropylethylamine (15.84 g, 122.8 mmol) was addeddropwise. Then, a solution of trifluoroacetic anhydride (8.25 g, 39.3mmol) in dichloromethane (20 ml) was added dropwise at −5° C. in thecourse of 30 minutes. After the mixture had been stirred for 30 minutesat 0° C., it was warmed to room temperature, and stirring was continuedfor another 12 hours. The mixture was diluted with dichloromethane (100ml) and washed with 20% strength citric acid, saturated NaHCO3 [sic]solution and saturated NaCl solution. The organic phase was dried(MgSO₄) and concentrated in vacuo. Yield: 6.3 g (25 mmol, quant.).

g) 4-Amidino-2-(N-Boc-aminomethyl)-5-methylthiazole×CH₃COOH

4-Cyano-2-(N-Boc-aminomethyl)-5-methylthiazole (5.5 g, 21.74 mmol) wasdissolved in methanol (15-ml), and N-acetylcysteine (4.1 g, 25.12 mmol)was added. The mixture was then warmed to 60° C., and ammonia was passedin for 22 hours. The batch was diluted with methanol and passed over anacetate ion exchanger. The methanol was removed on a rotary evaporatorand the residue extracted by stirring with acetone. The colorlessresidue was filtered off with suction and dried in vacuo. Yield: 4.75 g(14.4 mmol, 66%, colorless solid).

¹H NMR (400 MHz, DMSO-d₆)δ=1.40 (s, 9H), 1.80 (s, 3H), 2.60 (s, 3H),4.35 (d, J=6.5 Hz, 2H), 7.90 (t, J=6.5 Hz, 1H).

2-Aminomethyl-5-amidino-4-methylthiazole×2 HCl

a) N-BOC-Glycine [sic] Thioamide

N-BOC-Glycinonitrile [sic] (12.0 g, 76.8 mmol) and diethylamine (0.16ml, 2.1 mmol) were dissolved in toluene (100 ml). The solution wascooled to −10° C., saturated with hydrogen sulfide and subsequentlystirred overnight at room temperature. The precipitate formed wasfiltered off with suction and washed with toluene. The product was driedin vacuo at 45° C. Yield: 13.2 g (69.4 mmol, 90.3%, yellowish solid).

¹H NMR (270 MHz, DMSO-d₆) δ=1.40 (s, 9H), 3.80 (d, J=7 Hz, 2H), 7.05 (t,J=7 Hz, 1H), 9.0 (sbr, 1H), 9.65 (sbr, 1H).

b) Methyl 2-(N-BOC-Aminomethyl)-4-methylthiazole-5-carboxylate [sic]

N-BOC-Glycine [sic] thioamide (10.0 g, 52.6 mmol) was introduced intomethanol (70 ml), and methyl 2-chloroacetoacetate (7.9 g, 52.6 mmol) wasadded. The mixture was warmed for 2 hours at 60° C. and subsequentlystirred for 48 hours at room temperature. The methanol was removed on arotary evaporator and the residue was extracted by stirring withacetone/diethyl ether. The precipitate which remained was filtered offwith suction and the filtrate was concentrated. The solid obtained fromthe filtrate constituted the product (pure after TLC and HPLC). Yield:8.7 g (30.4 mmol, 57.8%). ESI-MS: 287 (M+H⁺).

2-(N-BOC-Aminomethyl)-4-methylthiazole-5-carboxylic [sic] Acid

Methyl 2-(N-BOC-aminomethyl)-4-methylthiazole-5-carboxylate [sic] (2.8g, 9.74 mmol) was dissolved in 1,4-dioxane (30 ml), and 1N sodiumhydroxide solution (19 ml) was added. After the mixture had been stirredfor 4 hours at room temperature, the 1,4-dioxane was removed on a rotaryevaporator. It was diluted with water and washed with ethyl acetate. Theaqueous phase was acidified with 20% strength potassium hydrogen sulfatesolution, and the precipitate obtained during this process was filteredoff with suction and washed with water. The product thus obtained wasdried in a vacuum drying oven at 40° C. Yield: 2.5 g.

d) 2-(N-BOC-Aminomethyl)-4-methylthiazole-5-carboxamide [sic]

2-(N-BOC-Aminomethyl)-4-methylthiazole-5-carboxylic [sic] acid (12.6 g,46.27 mmol) was dissolved in dichloromethane (460 ml) anddimethylformamide (0.4 ml). After the mixture had cooled to 0° C., asolution of oxalyl chloride (6.46 g, 50.90 mmol) in dichloromethane (40ml) was added dropwise in the course of 30 minutes. After the mixturehad been stirred for 2 hours at 0° C., it was cooled to −20° C., andammonia was passed in at this temperature until the reaction wascomplete. The mixture was subsequently warmed to room temperature andwashed with water. The precipitate formed during this process wasfiltered off with suction. The organic phase was washed with 5% strengthcitric acid solution, dried (MgSO₄) and concentrated on a rotaryevaporator. The resulting solid was combined with the precipitate whichhad previously been filtered off and dried at 50° C. in a vacuum dryingoven. Yield: 9.8 g (36.12 mmol, 78%).

e) 2-(N-BOC-Aminomethyl)-5-cyano-4-methylthiazole [sic]

2-(N-BOC-Aminomethyl)-4-methylthiazole-5-carboxamide [sic] (11.13 g,41.02 mmol) was suspended in dichloromethane (75 ml) and cooled to 0° C.At this temperature, ethyldiisopropylamine (17.86 ml, 102.55 mmol) wasfirst added, and then, slowly, a solution of trifluoroacetic anhydride(6.56 ml, 47.17 mmol) in dichloromethane (20 ml). After stirring for 1hour, the mixture was diluted with dichloromethane and washed with 5%strength citric acid solution. After drying (MgSO₄), the solvent wasremoved on a rotary evaporator and the crude product was purified byflash chromatography.

Yield: 6.5 g (25.66 mmol, 63%).

f) 2-(N-BOC-Aminomethyl)-4-methylthiazole-5-thioamide [sic]

2-(N-BOC-Aminomethyl)-5-cyano-4-methylthiazole [sic] (7.5 g, 29.61 mmol)was dissolved in pyridine (30 ml), and triethylamine (27 ml) was added.The solution was saturated with hydrogen sulfide at 0° C. and then leftto stand for 48 hours at room temperature. The solvent was subsequentlyremoved on a rotary evaporator, and the residue was taken up in ethylacetate, washed with 20% strength potassium hydrogen sulfate solutionand dried over magnesium sulfate. The solvent was removed on a rotaryevaporator, and the crude product was dissolved in dichloromethane andprecipitated with petroleum ether. The product which had precipitatedwas filtered off with suction and dried in a vacuum drying oven [lacuna]40° C. Yield: 7.1 g (24.7 mmol, 83%).

g) 5-Amidino-2-(N-BOC-aminomethyl)-4-methylthiazole [sic]×HOAc

2-(N-BOC-Aminomethyl)-4-methylthiazole-5-thioamide [sic] (7.1 g, 24.70mmol) was dissolved in dichloromethane (40 ml), and iodomethane (17.5 g,123.52 mmol) was added. After the mixture had been stirred for 56 hoursat room temperature, the solvent was removed on a rotary evaporator. Theresidue was dissolved in 10% strength methanolic ammonium acetatesolution (29 ml) and stirred at 40° C. until the reaction was complete.The solvent was removed on a rotary evaporator, the residue wasextracted by stirring with dichloromethane, and the resulting solid wasfiltered off with suction and washed with dichloromethane. The residuewas dissolved in methanol and converted into the corresponding acetateby means of an acetate-loaded ion exchanger. The solvent was removed ona rotary evaporator and the resulting reddish-brown oil was extracted bystirring with dichloromethane. During this process, the product wasobtained as colorless solid which was dried in vacuo at 400C. Yield: 5.3g (16.04 mmol, 65%).

h) 5-Amidino-2-aminomethyl-4-methylthiazole×2 HCl

5-Amidino-2-(N-BOC-aminomethyl)-4-methylthiazole [sic]×HOAC (1.6 g, 4.84mmol) was suspended in dichloromethane (20 ml), and 4M hydrochloric acidin 1,4-dioxane (4.84 ml, 19.37 mmol) was added at room temperature andthe mixture was stirred for 3 hours at this temperature. The product wasfiltered off, washed with dichloromethane and dried in vacuo at 40° C.

Yield: 0.73 g (3.00 mmol, 62%).

2-Aminomethyl-5-amidino-4-trifluoromethylthiazole×2 HCl

a) Ethyl 2-(N-BOC-Aminomethyl)-4-trifluoromethylthiazole-5-carboxylate[sic]

N-BOC-Glycine [sic] thioamide (5.0 g, 26.28 mmol) was dissolved inacetonitrile (60 ml), and a solution of ethyl2-chloro-4,4,4-trifluoroacetoacetate (6.38 g, 26.28 mmol) was addeddropwise at 5-10° C. Then, the mixture was stirred for a further 30minutes at 5° C. and for 12 hours at room temperature. The batch wasthen cooled to 0° C., and triethylamine (12 ml, 86.77 mmol) was addeddropwise. After the mixture had been stirred for 20 minutes at 0° C.,the yellow suspension had changed into a clear reddish-brown solution.Then, thionyl chloride (2.1 ml, 28.89 mmol) was slowly added dropwise at0° C. After the mixture had been stirred for 20 minutes at 0° C., it waswarmed to room temperature for a further hour. The solvent was removedon a rotary evaporator, and the residue was taken up in water (100 ml)and extracted repeatedly with ethyl acetate. The combined organic phaseswere dried (Na₂AO₄) and concentrated. The crude product was purified bychromatography (silica gel MeOH:DCM=2:98). Yield: 2.2 g (6.4 mmol,24.5%).

¹H NMR (270 MHz, DMSO-d₆) δ=1.30 (t, J=6.5 Hz, 3H), 1.45 (s, 9H), 4.35(q, J=6.5 Hz, 2H), 4.45 (d, J=6.5 Hz, 2H), 7.95 (t, J=6.5 Hz, 1H).

b) 2-(N-BOC-Aminomethyl)-4-tri-fluoromethylthiazole-5-carboxamide [sic]

Ethyl 2-(N-BOC-aminomethyl)-4-trifluoromethylthiazole-5-carboxylate[sic] (15 g, 42.33 mol) was dissolved in methanol. Ammonia was passedinto the solution at room temperature until all of the ester had beenconverted into the carboxamide. The solvent was removed on a rotaryevaporator and the crude product was purified by flash chromatography.Yield: 4.6 g (14.14 mmol, 33%).

c) 2-(N-BOC-Aminomethyl)-5-cyano-4-trifluoromethylthiazole [sic]

2-(N-BOC-Aminomethyl)-4-trifluoromethylthiazole-5-carboxamide [sic] (4.6g, 14.14 mmol) was dissolved in dichloromethane (30 ml) and cooled to−5° C. Ethyldiisopropylamine (4.6 g, 35.35 mmol) and a solution oftrifluoroacetic anhydride (3.4 g, 16.26 mmol) in dichloromethane (10 ml)were added at this temperature. Then, the mixture was stirred for afurther 2 hours at 0° C. It was washed in succession with saturatedsodium hydrogen carbonate solution and 5% strength citric acid solution.After drying (MgSO₄), the solvent was removed on a rotary evaporator.The crude product was extracted by stirring with diethyl ether/petroleumether. The supernatant was separated from the oil and concentrated on arotary evaporator. Yield: 1.9 g (6.18 mmol, 44%).

d) 2-(N-BOC-Aminomethyl)-4-trifluoromethylthiazole-5-thioamide [sic]

2-(N-BOC-Aminomethyl)-5-cyano-4-trifluoromethylthiazole [sic] (4.6 g,14.97 mmol) was dissolved in pyridine (20 ml), triethylamine (24 ml) wasadded, and the solution was saturated with hydrogen sulfide. After twodays at room temperature, the solvent was removed on a rotaryevaporator. The crude product was taken up in ethyl acetate and washedin succession with 20% strength sodium hydrogen sulfate solution andwater. After drying (MgSO₄), the solvent was removed on a rotaryevaporator. The crude product was purified by flash chromatography.Yield: 2.5 g (7.32 mmol, 49%).

e) 5-Amidino-2-(N-BOC-aminomethyl)-4-trifluoromethylthiazole [sic]

2-(N-BOC-Aminomethyl)-4-trifluoromethylthiazole-5-thioamide [sic] (2.5g, 7.32 mmol) was dissolved in dichloromethane (10 ml), and iodomethane(10.4 g, 73.24 mmol) was added. Then, the mixture was stirred for 48hours at room temperature. After the solvent had been removed on arotary evaporator, the residue was taken up in methanol (5 ml), and 10%strength methanolic ammonium acetate solution (8.5 ml, 10.98 mmol) wasadded. After the mixture had been stirred for 4 days at roomtemperature, the solution of the crude product was passed over anacetate-loaded ion exchanger and the solvent was removed on a rotaryevaporator. The crude product was purified by flash chromatography.Yield: 0.8 g (2.08 mmol, 28%).

f) 5-Amidino-2-aminomethyl-4-trifluoromethylthiazole×2 HCl

5-Amidino-2-(N-BOC-aminomethyl)-4-trifluoromethylthiazole [sic] (0.8 g,2.08 mmol) was dissolved in dichloromethane, and a 4M solution ofhydrochloric acid in 1,4-dioxane (2.1 ml, 4.2 mmol) was added. After themixture had been stirred for 1 hour at room temperature, the solvent wasremoved on a rotary evaporator. The crude product obtained in this waywas employed in the following reactions without further purification.Yield: 0.6 g (2.0 mmol, 97%). ESI-MS: 225 (M+H⁺).

5-Aminomethyl-3-methylthiophene-2-carbonitrile

a) 5-Formyl-3-methylthiophene-2-carbonitrile

112 ml (179 mmol) of a 1.6-molar solution of n-butyllithium in n-hexanewere added in the course of 20 minutes to a solution of 25.1 ml (179mmol) of diisopropylamine in 400 ml of tetrahydrofuran cooled to −78° C.The solution was allowed to come to −35° C., then cooled again to −78°C., and a solution of 20.0 g (162 mmol) of 2-cyano-3-methylthiophene in80 ml of tetrahydrofuran was slowly added dropwise at this temperature.During this process, the color of the solution changed to dark red.Stirring was continued for 45 minutes, 63 ml (811 mmol) ofdimethylformamide were slowly added dropwise, and the mixture wasstirred for another 30 minutes. For work-up, a solution of 27 g ofcitric acid in 160 ml of water was added at −70° C. The mixture wasconcentrated on a rotary evaporator, 540 ml of saturated sodium chloridesolution were added, and the batch was extracted three times using ineach case 250 ml of diethyl ether. The combined organic extracts weredried over magnesium sulfate. After the desiccant had been filtered off,the solvent was distilled off under a water pump vacuum and the residuewas purified by column chromatography (eluant hexane/ethyl acetate 4/1).This gave 23 g (94%) of the title compound. ¹H NMR (270 MHz, DMSO-d₆):δ=2.4 (s, 3H), 8.0 (s, 1H), 9.8 (s, 1H).

b) 5-Hydroxymethyl-3-methylthiophene-2-carbonitrile

5.75 g (152 mmol) of sodium borohydride were added portionwise at roomtemperature to a solution of 23 g (152 mmol) of5-formyl-3-methylthiophene-2-carbonitrile in 300 ml of absolute ethanol.The reaction mixture was stirred for 5 minutes, concentrated under awater pump vacuum, taken up in ethyl acetate and extracted with 5%strength citric acid solution and with saturated sodium chloridesolution, the organic phase was dried over magnesium sulfate, thedesiccant was filtered off, and the solvent was distilled off at roomtemperature and under a water pump vacuum. This gave 24 g of the titlecompound as a dark red oil which still contained solvent and which wasemployed in the following reactions without further purification. ¹H NMR(270 MHz, DMSO-d₆): δ=2.4 (s, 3H), 4.7 (m, 2H), 5.9 (m, 1H), 7.0 (s,1H).

c) 5-Bromomethyl-3-methylthiophene-2-carbonitrile

44 g (167 mmol) of triphenylphosphine were added to a solution of 24 g(152 mmol) of 5-hydroxymethyl-3-methylthiophene-2-carbonitrile in 180 mlof tetrahydrofuran. Then, a solution of 55 g (167 mmol) oftetrabromomethane in 100 ml of tetrahydrofuran was added. The mixturewas stirred for 90 minutes at room temperature. The reaction mixture wasconcentrated on a rotary evaporator under a water pump vacuum, and theresidue was purified by column chromatography (eluant hexane:ethylacetate 8:2). This gave 34 g of the title compound which still containeda small amount of solvent. ¹H NMR (270 MHz, DMSO-d₆): δ=2.4 (s, 3H), 5.0(s, 2H), 7.3 (s, 1H).

d)5-N,N-bis(tert-Butoxycarbonyl)aminomethyl-3-methylthiophene-2-carbonitrile

5.0 g (167 mmol) of sodium hydride (80% suspension in mineral oil) wasadded portionwise to a solution of 33.8 g (152 mmol) of5-bromomethyl-3-methylthiophene-2-carbonitrile in 255 ml oftetrahydrofuran, cooled to 0° C. Then, a solution of 36.4 g (167 mmol)of di-tert-butyl iminodicarboxylate in 255 ml of tetrahydrofuran wasadded dropwise, during this process the temperature did not rise above5° C. The mixture was allowed to come to room temperature and wasstirred overnight. To complete the reaction, the mixture was warmed fora further 3 hours at 35° C. and was then left to cool to roomtemperature, and 510 ml of a saturated ammonium chloride solution wasadded slowly. The solvent was distilled off under a water pump vacuum,the residue was extracted repeatedly with ethyl acetate, and thecombined organic phases were washed with saturated sodium chloridesolution, dried over magnesium sulfate and concentrated on a rotaryevaporator. This gave 57.6 g of an oily residue which still containeddi-tert-butyl iminodicarboxylate and which was employed in the followingreaction as a crude product. ¹H NMR (270 MHz, DMSO-d₆): δ=1.45 (s, 18H),2.35 (s, 3H), 4.85 (s, 2H), 7.05 (s, 1H).

e) 5-Aminomethyl-3-methylthiophene-2-carbonitrile Hydrochloride

52.6 g of5-N,N-bis(tert-butoxycarbonyl)aminomethyl-3-methylthiophene-2-carbonitrile(crude product of d), not more than 139 mmol) were dissolved in 950 mlof ethyl acetate and the solution was cooled to 0° C. It was saturatedwith hydrogen chloride gas, during which process a white precipitateseparated out after 10 minutes. The mixture was stirred for two hours atroom temperature and for one hour at 30° C., the resulting suspensionwas subsequently concentrated on a rotary evaporator, the residue wasextracted by stirring with diethyl ether, the solvent was filtered off,and the solid residue was dried at room temperature in vacuo. This gave24.7 g (94%) of the title compound as white powder.

¹H NMR (270 MHz, DMSO-d₆): δ=2.4 (s, 3H), 4.25 (s, 2H), 7.3 (s, 1H),8.8-9.0 (bs, 3H). ¹³CNMR (DMSO-d₆): 15.0 (CH3), 36.4 (CH₂), 104.8 (C-2),113.8 (CN), 131.5 (C-4), 142.8 (C-5), 149.6 (C-3).

5-Aminomethyl-3-chlorothiophene-2-carbonitrile Hydrochloride

This compound was synthesized as described for5-aminomethyl-3-methylthiophene-2-carbonitrile, the3-chloro-2-cyanothiophene employed having been prepared by dehydrating3-chlorothiophene-2-carboxamide with trifluoroacetic anhydride.

5-Aminomethyl-4-methylthiophene-3-thiocarboxamide

a) Ethyl 2-Amino-3-cyano-4-methylthiophene-5-carboxylate

Ethyl 2-amino-3-cyano-4-methylthiophene-5-carboxylate was synthesized asdescribed in “Organikum” [Organic Chemistry], 19th Edition, Dt. Verlagder Wissenschaften, Leipzig, Heidelberg, Berlin, 1993, Chapter 6,pp.374-375, starting from 130 g (1.0 mol) of ethyl acetoacetate, 66 g(1.0 mol) of malononitrile, 32 g (1.0 mol) of sulfur and 80 g (0.92 mol)of morpholin. ¹H NMR (270 MHz, DMSO-d₆): δ=1.25 (t, 3H), 2.3 (s, 3H),4.2 (q, 2H), 7.9 (bs, 2H).

b) Ethyl 4-Cyano-3-methylthiophene-2-carboxylate

A solution of 20.5 g (97.5 mmol) of ethyl2-amino-3-cyano-4-methylthiophene-5-carboxylate in 600 ml of a 1:1mixture of acetonitrile and dimethylformamide was cooled to 5° C., and15.7 g (146 mmol) of tert-butyl nitrite were added dropwise, duringwhich process the temperature of the reaction mixture rose and gas wasevolved vigorously. The mixture was stirred for seven hours at roomtemperature and concentrated on a rotary evaporator under a high vacuum,the residue was purified by column chromatography (eluantdichloromethane), and 9.1 g (48%) of the desired compound were obtainedas yellow oil. ¹H NMR (270 MHz, DMSO-d₆): δ=1.3 (t, 3H), 2.55 (s, 3H),4.3 (q, 2H), 8.8 (s, 1H).

c) 5-Hydroxymethyl-4-methylthiophene-3-carbonitrile

2.44 g (64 mmol) of lithium aluminum hydride were added portionwise at0° C. to a solution of 25.1 g (129 mmol) of ethyl3-cyano-4-methylthiophene-5-carboxylate in 400 ml of tetrahydrofuran.The mixture was stirred for 5 hours at room temperature, excess reducingagent was destroyed by adding 0.5 N hydrochloric acid, and the reactionmixture was concentrated under a water pump vacuum, diluted with waterand extracted three times with ethyl acetate. The combined organicphases were then washed in each case once with 0.5 N hydrochloric acidand saturated sodium chloride solution. The organic phase was dried overmagnesium sulfate, the desiccant was filtered off, and the solvent wasdistilled off under a water pump vacuum at room temperature. The residuewas purified by column chromatography (eluant dichloromethane/methanol95:5), and 16.1 g (83%) of the desired compound were obtained as aslightly yellow oil.

¹H NMR (270 MHz, DMSO-d₆): δ=2.2 (s, 3H), 4.6 (d, 2H), 5.7 (m, 1H), 8.35(s, 1H).

d) 5-Bromomethyl-4-methylthiophene-3-carbonitrile

30 g (115 mmol) of triphenylphosphine were added at 5° C. to a solutionof 16 g (104 mmol) of 5-hydroxymethyl-4-methylthiophene-3-carbonitrilein 300 ml of tetrahydrofuran. Then, a solution of 38 g (115 mmol) oftetrabromomethane in 100 ml of tetrahydrofuran was added. The mixturewas stirred overnight at room temperature. The reaction mixture wasconcentrated on a rotary evaporator under a water pump vacuum and theresidue was purified by column chromatography (eluant petroleumether:dichloromethane 1:1). This gave 17 g (76%) of the title compoundas yellow oil. ¹H NMR (270 MHz, DMSO-d₆): δ=2.25 (s, 3H), 5.0 (s, 2H),8.5 (s, 1H).

e)5-N,N-bis(tert-Butoxycarbonyl)aminomethyl-4-methylthiophene-3-carbonitrile

3.5 g (103 mmol) of sodium hydride (oil-free) was added portionwise to asolution of 17.2 g (79.5 mmol) of5-bromomethyl-4-methylthiophene-3-carbonitrile in 250 ml oftetrahydrofuran, cooled to 0° C. Then, a solution of 22.5 g (103 mmol)of di-tert-butyl iminodicarboxylate in 100 ml of tetrahydrofuran wasadded dropwise, during which process the temperature did not rise above5° C. The mixture was allowed to warm to room temperature and wasstirred for 2 hours. 400 ml of a saturated ammonium chloride solutionwas added slowly. The solvent was distilled off under a water pumpvacuum, and the residue was diluted with a little water and extractedthree times with ethyl acetate. The combined organic phases were washedwith saturated ammonium chloride solution and with saturated sodiumchloride solution, dried over magnesium sulfate and concentrated on arotary evaporator. This gave 28 g of an oil which still containeddi-tert-butyl iminodicarboxylate and was employed in the followingreaction as crude product. ¹H NMR (270 MHz, DMSO-d₆): δ=1.4 (s, 9H),1.45 (s, 9H), 2.3 (s, 3H), 4.8 (s, 2H), 8.4 (s, 1H).

f)5-N,N-bis(tert-Butoxycarbonyl)aminomethyl-4-methylthiophene-3-thiocarboxamide

The crude product obtained in e) (not more than 79 mmol) was dissolvedin 280 ml of pyridine and 140 ml of triethylamine and the solution wassaturated with hydrogen sulfide at room temperature. The color of thesolution, which was yellow at the beginning, changed to green. Themixture was stirred overnight at room temperature. To complete thereaction, hydrogen sulfide was passed in for a further 15 minutes andstirring was continued for two hours at room temperature. Excesshydrogen sulfide was expelled with the aid of a nitrogen stream using awashing tower. Then, the reaction mixture was concentrated on a rotaryevaporator, and the concentrate was taken up in ethyl acetate, washedrepeatedly with 20% strength sodium hydrogen sulfate solution, driedover magnesium sulfate and concentrated on a rotary evaporator. Thisgave 27 g of a pale yellow solid foam which wa s employed in thefollowing reaction without further purification. ¹H NMR (270 MHz,DMSO-d₆): δ=1.4 (s, 18H), 2.15 (s, 3H), 4.8 (s, 2H), 7.5 (s, 1H), 9.3(bs, 1H), 9.75 (bs, 1H).

g) 5-Aminomethyl-4-methylthiophene-3-thiocarboxamide Hydrochloride

27 g of5-N,N-bis(tert-butoxycarbonyl)-aminomethyl-4-methylthiophene-3-thiocarboxamide(crude product of f), not more than 70 mmol) were dissolved in 400 ml ofethyl acetate and the solution was cooled to 0° C. It was saturated withis hydrogen chloride gas, during which process a white precipitateseparated out after 10 minutes. The mixture was stirred for two hours atroom temperature, the precipitate was filtered off and washed with ethylacetate, and the solid residue was dried in vacuo at room temperature.This gave 13.6 g (87%) of the title compound as white powder. EI-MS:M⁺=186.

5-Aminomethyl-4-chlorothiophene-3-thiocarboxamide

a) 5-Formyl-4-chlorothiophene-3-carbonitrile

35 g (325 mmol) of tert-butyl nitrite were added dropwise at roomtemperature to a solution of 53.0 g (250 mmol) of2-amino-4-chloro-5-formylthiophene-3-carbonitrile (the synthesis of thiscompound is described in Patent DB 3738910) in 600 ml of a 1:1 mixtureof acetonitrile and dimethylformamide, during which process thetemperature of the reaction mixture rose from 20° C. to 37° C. andvigorous evolution of gas began. The mixture was cooled to 25° C. andstirred for 7 hours at room temperature, the black solution wasconcentrated on a rotary evaporator under a high vacuum, the residue waspurified by column chromatography (eluant dichloromethane), and 29 g(68%) of the desired compound were obtained as yellow oil. ¹H NMR (270MHz, DMSO-d₆): δ=9.1 (s, 1H), 10.0 (s, 1H).

b) 5-Hydroxymethyl-4-chlorothiophene-3-carbonitrile

6.3 g (166 mmol) of sodium borohydride were added portionwise at 5° C.to a solution of 28.5 g (166 mmol) of5-formyl-4-chlorothiophene-3-carbonitrile in 400 ml of rose slightly andthe color changed to dark red. A vigorous evolution of gas was observed.After 10 minutes, the reaction mixture was concentrated under a waterpump vacuum, taken up in 200 ml of ethyl acetate, extracted with 200 mlof 1 M hydrochloric acid and washed twice with in each case 250 ml ofwater and with saturated sodium chloride solution, the organic phase wasdried over magnesium sulfate, the desiccant was filtered off, and thesolvent was distilled off under a water pump vacuum at room temperature.This gave 22 g (76%) of the title compound as dark red oil which wasemployed in the following reactions without further purification. ¹H NMR(270 MHz, DMSO-d₆): δ=4.65 (bs, 1H), 5.95 (t, 2H), 8.6 (s, 1H).

c) 5-Bromomethyl-4-chlorothiophene-3-carbonitrile

36.1 g (137 mmol) of triphenylphosphine were added at 5° C. to asolution of 21.7 g (125 mmol) of5-hydroxymethyl-4-chlorothiophene-3-carbonitrile in 250 ml oftetrahydrofuran. Then, a solution of 45.6 g (137 mmol) oftetrabromomethane in 100 ml of tetrahydrofuran was added. The mixturewas stirred overnight at room temperature. The precipitate was filteredoff, the filtrate was concentrated on a rotary evaporator under a waterpump vacuum, and the residue was purified by column chromatography(eluant petroleum ether:dichloromethane 1:1). This gave 26.0 g (88%) ofthe title compound as an oil.

¹H NMR (270 MHz, DMSO-d₆): δ=4.95 (s, 2H), 8.8 (s, 1H).

d)5-N,N-bis(tert-Butoxycarbonyl)aminomethyl-4-chlorothiophene-3-carbonitrile

6.9 g (159 mmol) of sodium hydride (oil-free) was added portionwise to asolution of 25.0 g (106 mmol) of5-bromomethyl-4-chlorothiophene-3-carbonitrile in 300 ml oftetrahydrofuran, cooled to 0° C. Then, a solution of 34.4 g (159 mmol)of di-tert-butyl-iminodicarboxylate in 100 ml of tetrahydrofuran wasadded dropwise, during which process the temperature did not rise above5° C. The mixture was allowed to warm to room temperature and wasstirred for two hours. 300 ml of a saturated ammonium chloride solutionwere added slowly. The solvent was distilled off under a water pumpvacuum, and the residue was diluted with a little water and extractedthree times with ethyl acetate. The combined organic phases were washedwith saturated ammonium chloride solution and with saturated sodiumchloride solution, dried over magnesium sulfate and concentrated on arotary evaporator. This gave 51.3 g of an oil which still containeddi-tert-butyl iminodicarboxylate and solvent residues and which wasemployed in the following reaction as crude product. ¹H NMR (270 MHz,DMSO-d₆): δ=1.4 (s, 9H), 1.45 (s, 9H), 4.8 (s, 2H), 8.65 (s, 1H).

e)5-N,N-bis(tert-Butoxycarbonyl)aminomethyl-4-methylthiophene-3-thiocarboxamide

Some of the crude product obtained in d) (39.4 g, not more than 106mmol) was dissolved in 400 ml of pyridine and 40 ml of triethylamine andthe solution was saturated with hydrogen sulfide at room temperature.The color of the solution, which was yellow at the beginning, changed togreen. The mixture was stirred overnight at room temperature. Excesshydrogen sulfide was expelled with the aid of a stream of nitrogen usinga washing tower. The reaction mixture was then poured into ice-cooled20% strength sodium hydrogen sulfate solution and extracted three timeswith ethyl acetate. The organic phase was then washed repeatedly with20% strength sodium hydrogen sulfate solution, dried over magnesiumsulfate and concentrated on a rotary evaporator. This gave 49.0 g of asolvent-containing residue which was employed in the following reactionwithout further purification. ¹H NMR (270 MHz, DMSO-d₆): δ=1.4, 1.45 (s,18H), 4.8 (s, 2H), 7.75 (s, 1H), 9.4 (bs, 1H), 10.0 (bs, 1H).

f) 5-Aminomethyl-4-chlorothiophene-3-thiocarboxamide Hydrochloride

38.0 g of the crude product of e), not more than 93 mmol, were dissolvedin 400 ml of ethyl acetate and cooled to 0° C. The solution wassaturated with hydrogen chloride gas, during which process a whiteprecipitate separated out after 10 minutes. Since the reaction was stillincomplete, 200 ml of ethyl acetate were added, and the mixture wassaturated again with hydrogen chloride gas and stirred overnight at roomtemperature. The precipitate was filtered off, washed with petroleumether and dried in vacuo at room temperature. This gave 21.1 g of thetitle compound as white powder which contained ammonium chloride ascontamination. EI-MS: M⁺=206.

5-Aminomethyl-1-methyl-1H-[1,2,4]-triazole-3-carboxamide

a) Ethyl Aminothioxoacetate

Hydrogen sulfide was passed to saturation at 0° C. in a solution of 29.1g (294 mmol) of ethyl cyanoformate and 0.4 g (0.57 ml, 5.1 mmol) ofdiethylamine in 20 ml of benzene, during which process the solutionturned orange. The mixture was stirred over the weekend at roomtemperature, the reaction mixture was cooled to 0° C., and theprecipitate formed (29.1 g) was filtered off and washed with coldbenzene. The mother liquor was concentrated and again cooled to 0° C.The mixture was filtered off, the residue was washed with petroleumether, and a further 5.7 g of the title compound were obtained as paleyellowish solid (Rf=0.7, dichloromethane/methanol 9:1). Overall yield:89%. ¹H NMR (270 MHz, DMSO-d6): δ=1.25 (t, J=7 Hz, 3H), 4.2 (q, J=7 Hz,2H) 9.9 (bs, 1H, NH), 10.4 (bs, 1H, NH).

b) Ethyl Methyloxamidrazonecarboxylate

A solution of 11.93 g (13.6 ml, 259 mmol) of methylhydrazine in 100 mlof ethanol was added at room temperature dropwise to a solution of 34.5g (259 mmol) of ethyl aminothioxoacetate in 400 ml of ethanol, duringwhich process the temperature of the reaction mixture rose slightly. Themixture was stirred for three hours at room temperature andconcentrated, and the residue was employed in reaction c) withoutfurther purification.

c) EthylAmino[(2-tert-butoxycarbonylaminoacetyl)methyl]-hydrazonoacetate

Activation of Boc-Gly-OH and reaction with b):

37.7 g (51.7 ml, 373 mmol) of triethylamine were added at roomtemperature to a solution of 54.46 g (311 mmol) of Boc-glycine in 400 mlof tetrahydrofuran. The mixture was cooled to −5° C., and a solution of40.47 g (35.5 ml, 311 mmol) of ethyl chloroformate in 100 ml oftetrahydrofuran was slowly added dropwise in the course of 40 minutes.The mixture was stirred for 30 minutes at −5° C., the resultingprecipitate was filtered off and washed with a small amount oftetrahydrofuran, and the filtrate was directly reacted further by slowlyadding dropwise at room temperature a solution of the residue from b)(259 mmol) in 300 ml of tetrahydrofuran. The mixture was stirredovernight and concentrated to dryness on a rotary evaporator underreduced pressure, and the residue was purified by column chromatography(silica gel, dichloromethane/methanol 95:5, Rf=0.26). This gave 15.7 gof an oil, which was taken up in diethyl ether, and the precipitate wasfiltered off (8.5 g, 11%). ¹H NMR (270 MHz, DMSO-d6): δ=1.25 (t, J=7 Hz,3H), 1.35 (s, 9H), 2.9 (s, 3H), 3.6 (d, J=5 Hz, 2H), 4.3 (q, J=7 Hz, 2H)6.6 (t, J=5 Hz 1H), 7.3 (bs, 2H).

d) Ethyl 5-Aminomethyl-1-methyl-1H-[1,2,4]-triazole-3-carboxylate

7.0 g (23.2 mmol) of ethylamino[(2-tert-butoxycarbonylaminoacetyl)methyl]hydrazonoacetate weresuspended in 30 ml of xylene and the suspension was immersed for 10minutes in a silicone oil bath which had been preheated to 180° C. Then,the solvent was distilled off directly from the reaction mixture and theresidue was stirred for a further 10 minutes at 180° C. Solvent residueswere removed at 50° C. under a high vacuum, and 6.8 g (>95%) of a darkoil were obtained which oil was employed in the following reactionwithout further purification. A sample was filtered through silica geland examined by NMR spectroscopy. ¹H NMR (270 MHz, DMSO-d6): δ=1.25 (t,J=7 Hz, 3H), 1.35 (s, 9H), 3.9 (s, 3H), 4.2-4.4 (m, 4H), 7.5 (t, J=5 Hz,1H).

e) 5-Aminomethyl-1-methyl-1H-[1,2,4]-triazole-3-carboxamide

Ammonia gas was passed for 20 minutes at −10° C. into a solution of 6.8g (not more than 23.2 mmol) of ethyl5-aminomethyl-1-methyl-1H-[1,2,4]-triazole-3-carboxylate in 200 ml ofethanol. Stirring was continued for one hour at 0° C. and overnight atroom temperature. Since the reaction was incomplete, the procedure ofpassing in gas was repeated twice more (as described above) and themixture was stirred overnight at 0° C. The mixture was concentrated on arotary evaporator and the residue was purified by column chromatography(dichloromethane +5-10% methanol, Rf=0.3 in dichloromethane/methanol9:1). This gave 4.71 g as colorless oil. ¹H NMR (270 MHz, DMSO-d6):δ=1.4 (s, 9H), 3.85 (s, 3H), 4.3 (d, J=5 Hz, 3H), 7.4 (bs, 1H), 7.6 (bs,1H), 7.65 (bs, J=5 Hz, 1H).

f) 5-Aminomethyl-1-methyl-1H-[1,2,4]-triazole-3-carboxamideHydrochloride

Hydrogen chloride was passed to saturation at 5° C. to a solution of 4.7g (not more-than 18.4 mmol) of5-aminomethyl-1-methyl-1H-[1,2,4]-triazole-3-carboxamide in 600 ml ofethyl acetate, during which process a white precipitate formed. Themixture was stirred overnight at room temperature and concentrated on arotary evaporator, diethyl ether was added, the mixture was concentratedand again taken up in diethyl ether, and the precipitate was filteredoff and dried. This gave 3.7 g of a white solid which still containedammonium chloride. ¹H NMR (270 MHz, DMSO-d6): δ=3.95 (s, 3H), 4.3 (bs,2H), 7.6 (bs, 1H), 7.75 (bs, 1H), 8.7-8.9 (m, 2H).

5-Aminomethyl-3-cyanofuran Hydrochloride

a) 5-N,N-bis(tert-Butoxycarbonyl)aminomethyl-3-cyanofuran

A solution, cooled to 0° C., of 20.5 g (0.11 mol) of5-bromomethyl-3-cyanofuran (L. M. Pevzner, V. M. Ignat'ev, B. I. Ionin,Russ. J. of Gen. Chem. 1994, 64, 2, 125-128) in 50 ml of tetrahydrofuranwas added with stirring in the course of 30 minutes at 0° C. to asuspension of 4.8 g (0.12 mol) of sodium hydride (60% dispersion inmineral oil) in 30 ml of tetrahydrofuran. A solution of 26.2 g (121mmol) of di-tert-butyl iminodicarboxylate in 50 ml of tetrahydrofuranwas subsequently added dropwise, during which process the temperaturedid not climb above 5° C. The mixture was stirred for three hours at5-10° C., allowed to warm to room temperature and stirred overnight. 150ml of a saturated ammonium chloride solution were slowly added. Thesolvent was distilled out under a water pump vacuum, the residue wasextracted four times using in each case 60 ml of ethyl acetate, and thecombined organic phases were washed twice using saturated sodiumchloride solution, dried over magnesium sulfate and concentrated in arotary evaporator. After drying for three hours at room temperature invacuo (1 mm Hg), 33.2 g of a dark syrup which still containeddi-tert-butyliminodicarboxylate resulted, and this was employed as crudeproduct in the reaction below. ¹H NMR (250 MHz, d₆-DMSO): δ=1.40, 1.45(s, 18H), 4.70 (s, 2H), 6.70 (s, 1H), 8.6 (s, 1H).

b) 5-Aminomethyl-3-cyanofuran Hydrochloride

12.89 g of 5-N,N-bis(tert-butoxycarbonyl)aminomethyl-3-cyanofuran (crudeproduct from a) were dissolved in 80 ml of ethyl acetate and cooled to−10° C. The mixture was saturated with hydrogen chloride gas, a whiteprecipitate separating out after 15 minutes. The mixture was allowed tocome to room temperature and was stirred for two hours, the resultingsuspension was subsequently concentrated in a rotary evaporator, theresidue (7 g) was extracted by stirring with diethyl ether, solvent wasremoved by filtration, and the solid residue was dried in vacuo at roomtemperature. 5 g (79%) of the title compound resulted as a pale ochrepowder. ¹H NMR (250 MHz, d₆-DMSO): δ=4.15 (bs, 2H), 7.0 (s, 1H), 8.6-8.9(m, 4H).

5-Aminomethyl-1-methylpyrrole-2-carbonitrile

a) 5-Cyano-1-methylpyrrole-2-carbaldehyde

1-Methylpyrrole was converted into 2-cyano-1-methylpyrrole by reactionwith chlorosulfonyl isocyanate and dimethylformamide in acetonitrile(see, for example, C. E. Loader et al. Can. J. Chem. (1981), 59,2673-6).

Diisopropylamine (17.5 ml, 124.38 mmol) was introduced into THF (100 ml)under nitrogen. N-Butyllithium solution in hexane (15% strength, 75.9ml, 124.38 mmol) was added dropwise at −78° C. The mixture wassubsequently stirred for 45 minutes at −20° C. and then cooled again to−78° C. At this temperature, a solution of1-methylpyrrole-2-carbonitrile (12 g, 113.07 mmol) in THF (50 ml) wasadded dropwise. After stirring for 45 minutes at −78° C., DMF (43.9 ml,546.46 mmol) was added dropwise, and the mixture was stirred at thistemperature for a further 2 hours. After addition of citric acidmonohydrate (20.56 g), the mixture was warmed to room temperature, andwater (112 ml) was added. The THF was removed in a rotary evaporator,and the aqueous phase was saturated with sodium chloride and extractedwith diethyl ether (3×200 ml). The combined organic phases were washedwith saturated sodium chloride solution and dried over sodium sulfate.The solvent was removed in a rotary evaporator and the crude product waspurified by means of flash chromatography (silica gel, dichloromethane).Yield: 8.25 g (54%).

1H [sic] NMR (CDCl₃) δ=4.1 (s, 3H), 6.8 (d, 1H), 6.9 (d, 1H), 9.7 (s,1H).

b) 5-Hydroxymethyl-1-methylpyrrole-2-carbonitrile

The product obtained in accordance with a) (8.2 g, 61.1 mmol) wasdissolved in ethanol (200 ml), and sodium borohydride (2.31 g, 61.13mmol) was added at −10° C. After stirring for 1.5 hours at 0-5° C., thesolvent was removed in a rotary evaporator, and ice-water and 20%strength sodium hydrogen sulfate solution were added to the residue. Theaqueous phase was extracted with ethyl acetate. The combined organicphases were washed to neutrality with saturated sodium hydrogencarbonate solution and water and dried over sodium sulfate. The solventwas removed in a rotary evaporator and the crude product was purified bymeans of flash chromatography (silica gel,dichloromethane/methanol=97.5/2.5). Yield: 7.6 g (91%). 1H [sic] NMR(CDCl₃) δ=1.9 (t, 1H), 3.75 (s, 3H), 4.6 (d, 2H), 6.1 (d, 1H), 6.7 (d,1H).

c) 5-Azidomethyl-1-methylpyrrole-2-carbonitrile

The product obtained in accordance with b) (7.5 g, 55.08 mmol) wasdissolved in DMF (220 ml), and triphenylphosphine (43.34 g, 165.25 mmol)was added at 0° C. After stirring for 5 minutes at this temperature,tetrabromomethane (54.8 g, 165.25 mmol) was added. The mixture wassubsequently stirred for 30 minutes at 0° C. and for 1.5 hours at roomtemperature. After cooling to 0° C., sodium azide (4.37 g, 67.21 mmol)was added. The mixture was subsequently stirred for 4.5 hours at roomtemperature. Saturated sodium chloride solution was added dropwise at 0°C., and the batch was diluted with ethyl acetate. The organic phase wasseparated off, and the aqueous phase was extracted with diethyl ether.The combined organic phases were washed with water and dried over sodiumsulfate. The solvent was removed in a rotary evaporator and the crudeproduct was purified by means of flash chromatography (silica gel, ethylacetate/hexane=1/20).

Yield: 5.6 g (63%). 1H [sic] NMR (CDCl₃) δ=3.75 (s, 3H), 4.35 (s, 2H),6.2 (d, 1H), 6.7 (d, 1H).

d) 5-Aminomethyl-1-methylpyrrole-2-carbonitrile

The product obtained in accordance with c) (4.71 g, 29.25 mmol) wasdissolved in methanol (100 ml), and palladium on charcoal (10%, 1 g) wasadded. The mixture was subsequently hydrogenated with hydrogen under 1atmosphere for 4 hours. The catalyst was removed by filtration throughCelitee and the filtrate was evaporated in a rotary evaporator. Theresidue was extracted by stirring with dichloromethane/diethylether=1/1. The product was filtered off with suction and dried at 35° C.in a vacuum drying oven.

Yield: 2.7 g (68%). 1H [sic] NMR (CDCl₃) δ=3.75 (s, 3H), 3.85 (s, 2H),6.05 (d, 1H), 6.7 (d, 1H).

Aminomethyl-1-methylpyrrole-2-carbonitrile

a) 5-Cyano-1-methylpyrrole-3-carbaldehyde

Aluminum trichloride (24.24 g, 180.86 mmol) was dissolved innitromethane/dichloromethane (1/1, 320 ml), the solution was cooled to−20° C., and 1-methylpyrrole-2-carbonitrile (8 g, 75.36 mmol) was added.α,α-Dichlorodimethyl ether (10.4 g, 90.43 mmol), dissolved indichloromethane (42 ml), was subsequently added dropwise. After stirringfor 4 h at 0° C., the batch was poured onto ice (200 g). The aqueousphase was extracted with diethyl ether. The combined organic phases werewashed until neutral with saturated sodium hydrogen carbonate solution,water and saturated sodium chloride solution. After drying over sodiumsulfate, the solvent was removed in a rotary evaporator. The crudeproduct was employed in the reactions below without furtherpurification.

Yield: 9.2 g (91%). 1H [sic] NMR (CDCl₃) δ=3.8 (s, 3H); 7.2 (s, 1H); 7.4(s, 1H); 9.85 (s, 1H).

b) Starting from 5-Cyano-1-methylpyrrole-3-carbaldehyde;4-aminomethyl-1-methylpyrrole-2-carbonitrile was synthesized analogouslyto the synthesis of 5-aminomethyl-1-methylpyrrole-2-carbonitrile.However, the 4-azidomethyl-1-methylpyrrole-2-carbonitrile wasadvantageously reduced in a Staudinger reaction (see S. Nagarajan et al.J. Org. Chem. 1987, 52, 5044-6).

1H [sic) NMR (DMSO-d₆) δ=3.77 (s, 3H), 3.84 (sbr, 2H), 7.00 (sbr, 1H),7.26 (s, 1H), 8.05 (sbr, 2H).

5-Aminomethyl-1-methylpyrrole-3-carbonitrile

a) 4-Cyano-1-methylpyrrole-2-carbaldehyde

1-Methylpyrrole-2-carbaldehyde (10 g, 91.6 mmol) was dissolved inacetonitrile (100 ml) and cooled to −45° C. Chlorosulfonyl isocyanate(38.9 g, 274.9 mmol) in acetonitrile (40 ml) was added dropwise in thecourse of 40 minutes. The mixture was subsequently stirred for 12 hoursat room temperature. After dropwise addition of dimethylformamide (35ml), the mixture was warmed to 50° C. for 1 hour. After cooling to roomtemperature, the reaction mixture was poured onto ice (200 ml) and 2Nsodium hydroxide solution (286 ml). The precipitate formed was filteredoff with suction. The filtrate was extracted with diethyl ether. Thecombined ether phases were washed until neutral with dilute sodiumhydrogen carbonate solution and water and dried over sodium sulfate. Thesolvent was distilled out in a water pump vacuum and the residue wascombined with the precipitate previously obtained. Recrystallizationfrom petroleum ether gave 4-cyano-1-methylpyrrole-2-carbaldehyde (4.3 g)(see, for example, C. E. Loader et al. Can. J. Chem. (1981), 59, 2673-6)1-H [sic] NMR (CDCl₃) δ=4.0 (s, 3H); 7.2 (s, 1H); 7.3 (s, 1H); 9.6 (s,1H).

13-C [sic] NMR (CDCl₃) δ=37.4; 94.1; 114.7; 125.8; 132.2; 135.8; 179.7.

b) Starting From 4-Cyano-1-methylpyrrole-2-carbaldehyde,5-Aminomethyl-1-methylpyrrole-3-carbonitrile was Prepared Analogously tothe Synthesis of 5-Aminomethyl-1-methylpyrrole-2-carbonitrile.

1H [sic] NMR (DMSO-d₆) δ=3.6 (s, 3H), 3.8. (s, 2H), 4.2 (sbr, 2H), 6.4(s, 1H), 7.6 (s, 1H).

5-Aminomethyl-3-cyano-1,2,4-oxadiazole hydrochloride

a) N-Boc-5-Aminomethyl-3-cyano-1,2,4-oxadiazole

Ethyl N-Boc-5-aminomethyl-1,2,4-oxadiazole-2-carboxylate (S. Borg et al.J. Org. Chem. 1995, 60, 3112-20) was dissolved in methanol (50 ml).Ammonia was passed into this solution at −10° C. to RT until thereaction was complete. The solvent was removed in a rotary evaporator.The resulting crude product was dissolved in dichloromethane (70 ml),and diisopropylethylamine (2.9 ml, 16.55 mmol) was added at −5° C.Trifluoroacetic anhydride (1.06 ml, 7.61 mmol), dissolved indichloromethane (10 ml), was subsequently added dropwise. After stirringfor 1.5 hours at 0° C., the batch was diluted with dichloromethane,washed 2× with saturated sodium hydrogen carbonate solution, 2× with 5%strength citric acid solution and 1× with saturated sodium chloridesolution and then dried over sodium sulfate. The solvent was removed ina rotary evaporator and the crude product was purified by chromatography(silica gel, dichloromethane:methanol=97.5:2.5). Yield: 1.2 g (80%).

b) 5-Aminomethyl-3-cyano-1,2,4-oxadiazole Hydrochloride

The product obtained in accordance with a) (0.9 g, 4.0 mmol) wasdissolved in dichloromethane (45 ml), and 4M hydrochloric acid indioxane (3.9 ml, 15.61 mmol) was added at RT. After stirring for 16hours at RT, the solvent was removed in a rotary evaporator. Yield: 645mg (100%).

1-H [sic] NMR (DMSO-d₆) δ=4.6 (s, 2H), 9.2 (s, 3H).

1-Methyl-5-aminomethylpyrazole-3-carboxamide

a) Methyl 1-Methyl-5-amidopyrazole-3-carboxylate

1-Methyl-3-methoxycarbonylpyrazole-5-carboxylic acid chloride (preparedfrom 3.7 g, 20.09 mmol, of 1-methyl-3-methoxycarbonyl-3-carboxylic acid,J. Org. Chem. 1989, 54, 428) was dissolved in toluene and the solutionwas cooled to −10° C. Ammonia was subsequently passed in at −10° C. to0° C. until the reaction was complete. The solvent was removed in arotary evaporator. The residue was taken up in ethanol. After stirringfor 15 minutes, the ethanol was removed in a rotary evaporator, and theresidue was dissolved in warm water and precipitated by cooling thesolution to 0° C. The precipitate was filtered off with suction, washedwith acetone and dried in vacuo at 45° C. Yield: 1.5 g (41%).

b) Methyl 1-Methyl-5-cyanopyrazole-3-carboxylate

The product obtained in accordance with a) (1.5 g, 8.19 mmol) were [sic]taken up in dichloromethane (20 ml). Diisopropylethyl- amine (3.85 ml,22.11 mmol) was added at −10° C., and a solution of trifluoroaceticanhydride (1.3 ml, 9.44 mmol) in dichloromethane (5 ml) was addeddropwise at this temperature in the course of 45 minutes. Stirring wassubsequently continued for 1 hour at 0° C. The batch was diluted withdichloromethane and washed 2× with saturated sodium hydrogen carbonatesolution, 2× with 5% strength citric acid solution and 1× with saturatedsodium chloride solution. After drying over sodium sulfate, the solventwas removed in a rotary evaporator. Yield: 1.35 g (100%).

c) 1-Methyl-5-cyanopyrazole-3-carboxamide

The product obtained in accordance with b) (1.35 g, 8.19 mmol) wasintroduced into methanol (50 ml) and cooled to −10° C. Ammonia wassubsequently passed in in the course of 8 hours. After stirring for 12hours at room temperature, reaction of the precursor had ended. Theproduct which had precipitated was filtered off with suction, washedwith cold methanol and dried in vacuo. Yield: 1.22 g (100%).

1-H [sic] NMR (DMSO-d₆) δ=4.0 (s, 3H), 7.4 (s, 1H), 7.5 (s, 1H), 7.8 (s,1H).

d) 1-Methyl-5-aminomethylpyrazole-3-carboxamide

The product obtained in accordance with c) (0.4 g, 2.66 mmol) wasdissolved in acetic acid (30 ml) and 10% palladium on charcoal (78 mg)was added. The mixture was subsequently hydrogenated at room temperatureunder atmospheric pressure until the reaction was complete. The catalystwas removed by filtration through Celite® and the solvent was removed ina rotary evaporator. Yield: 0.4 g (100%), FAB-MS (M+H⁺): 155.

1-Methyl-3-aminomethyl-pyrazole-5-carboxamide

a) Methyl 1-Methyl-3-amidopyrazole-5-carboxylate

1-Methyl-5-methoxycarbonylpyrazole-3-carbonyl chloride (synthesized from4.17 g, 22.6 mmol, of 1-methyl-5-methoxycarbonyl-3-carboxylic acid, J.Org. Chem. 1989, 54, 428) was dissolved in toluene and the solution wascooled to −10° C. Then, ammonia was passed in at −10° C. to 0° C. untilthe reaction was complete. The solvent was removed in a rotaryevaporator. The residue was taken up in ethanol. After the mixture hadbeen stirred for 15 minutes, the ethanol was removed in a rotaryevaporator, and the residue was dissolved in warm water and precipitatedby cooling to 0° C. The precipitate was filtered off with suction,washed with acetone and dried in vacuo at 45° C. Yield: 3.36 g (18.4mmol, 81%).

¹H NMR (270 MHz, DMSO-d₆) δ=3.85 (s, 3H), 4.15 (s, 3H), 7.20 (s, 1H),7.4 (sbr, 1H), 7.7 (sbr, 1H).

b) Methyl 1-Methyl-3-cyanopyrazole-5-carboxylate

The product obtained in a) (3.36 g, 18.4 mmol) was reacted similarly tothe method described above for the synthesis of methyl1-methyl-cyanopyrazole-3-carboxylate. Yield: 2.59 g (15.7 mmol, 85%).

¹H NMR (250 MHz, DMSO-d₆) δ=3.90 (s, 3H), 4.15 (s, 3H), 7.60 (s, 1H).

c) 1-Methyl-3-cyanopyrazole-5-carboxamide

The product obtained in b) (2.56 g, 15.5 mmol) was reacted similarly tothe method described above for the synthesis of1-methyl-5-cyanopyrazole-3-carboxamide. Yield: 2.3 g (15.3 mmol, 99%).

¹H NMR (250 MHz, DMSO-d₆) δ=4.15 (s, 3H), 7.45 (s, 1H), 7.70 (sbr, 1H),8.15 (sbr, 1H).

d) 1-Methyl-3-aminomethylpyrazole-5-carboxamide×HCl

The product obtained in c) (1.0 g, 6.7 mmol) was reacted similarly tothe method described above for the synthesis of1-methyl-5-aminomethylpyrazole-3-carboxamide. Yield: 1.5 g (5.6 mmol,83%).

¹H NMR (270 MHz, DMSO-d₆) δ=4.00 (q, J=6.5 Hz, 2H), 4.10 (s, 3H), 6.90(s, 1H), 7.60 (sbr, 1H), 8.05 (sbr, 1H), 8.25 (sbr, 3H).

The product can be converted into the corresponding hydrochloride byrepeatedly treating it with HCl in 1,4-dioxane and subsequentlyconcentrating the mixture.

EXAMPLE 1N-(Hydroxycarbonylmethylene)-(D)-cyclohexylalanylprolyl-[2-(4-amidino)thiazolylmethyl]amideHydrochloride

a) N-(t-BuO₂C—CH₂)-N-Boc-(D)-Cha-Pro-NH—CH₂-2-(4-CSNH₂)-thiaz

N-(t-BuO₂C—CH₂)-N-Boc-(D)-Cha-Pro-OH (2.0 g, 4.14 mmol),2-H₂N—CH₂-thiaz-4-CSNH₂ (1.0 g, 4.56 mmol) and diisopropylethylamine(5.5 ml, 32.53 mmol) were dissolved in 25 ml of methylene chloride, thesolution was cooled to 0° C., and 4.8 ml (6.21 mmol) of a 50% strengthsolution of propanephosphonic anhydride in ethyl acetate was addeddropwise. The reaction mixture was stirred for 1 hour at 0° C. and for 1hour at room temperature and subsequently concentrated in vacuo, theresidue was taken up in water, the mixture was extracted repeatedly withether, and the organic phase was dried over magnesium sulfate andconcentrated in vacuo. Because of slight impurities, the product waspurified by chromatography over silica gel. The pure fractions werecrystallized from ether. This resulted in a total of 1.9 g of therequired product.

b) N-(t-BuO₂C—CH₂)-N-Boc-(D)-Cha-Pro-NH—CH₂-2-(4-C(SCH₃)NH)-ThiazHydroiodide

N-(t-BuO₂C—CH₂)-N-Boc-(D)-Cha-Pro-NH—CH₂-2-(4-CSNH₂)-thiaz (1.7 g, 2.67mmol) together with 3.7 ml of methyl iodide in 30 ml of methylenechloride were [sic] stirred overnight at room temperature, subsequentlyconcentrated in vacuo under mild conditions and employed in thesubsequent reaction in the form of the crude product (2.08 g, max 2.67mmmol [sic]).

c) N-(t-BuO₂C—H₂)-N-Boc-(D)-Cha-Pro-NH—CH₂-2-(4-C(NH₂)NH)-ThiazHydroacetate

N-(t-BuO₂C—CH₂)-N-Boc-(D)-Cha-Pro-NH-H₂-2-(4-C(SCH₃)NH)-thiazhydroiodide (2.08 g, max 2.67 mmol) were [sic] dissolved in 20 ml ofacetonitrile, 0.6 g (8.01 mmol) of ammonium acetate was added, and themixture was stirred for 1.5 hours at 40-50° C. After the solvent hadbeen evaporated in vacuo in a rotary evaporator, the residue was takenup in methylene chloride, insoluble excess ammonium acetate was removedby filtration, the methylene chloride solution was concentrated, theresidue was taken up in ether, and the required product was precipitatedwith n-hexane as an amorphous solid substance. The crude product (2.1 g)was dissolved in 20 ml of methanol and converted into the correspondingacetate by means of acetate ion exchanger (3.7 g, Fluka, Product No.00402).

d) HOOC—CH₂-(D)-Cha-Pro-NH—CH₂-2-(4-am)-Thiaz Dihydrochloride

N-(t-BUO₂C_CH₂)-N-Boc-(D)-Cha-Pro-NH—CH₂-2-(4-C(NH₂)NH)-thiaz×CH₃COOH(2.0 g, max 2.67 mmol) were [sic] heated for 4 hours at 40-50° C. in amixture of 10 ml of dioxane and 20 ml of 5N aqueous hydrochloric acidsolution, the mixture was subsequently extracted repeatedly withmethylene chloride, and the aqueous phase was slightly concentrated invacuo and subsequently freeze-dried. 1.4 g ofHOOC—CH₂-(D)-Cha-Pro-NH—CH₂-2-(4-am)-thiaz dihydrochloride were obtainedas a white amorphous solid substance, FAB-MS (M+H⁺): 465.

EXAMPLE 2N-(Hydroxycarbonylmethylene)-(D)-cyclohexylalanylprolyl-[4-(2-amidino)thienylmethyl]amideHydroacetate

a) N-(t-BuO₂C—CH₂)-N-Boc-(D)-Cha-Pro-NH—CH₂-4-(2-CN)-Thioph

Starting from N-(t-BuO₂C—CH₂)-N-Boc-(D)-Cha-Pro-OH (6.35 g, 13.17 mmol)and 4-H₂N—CH₂-thioph-2-CN (2.3 g, 13.17 mmol), coupling was effectedanalogously to Example 1 to giveN-(t-BuO₂C—CH₂)-N-Boc-(D)-Cha-Pro-NH—CH₂-4-(2-CN)-thioph, 6.95 g of therequired product resulting after purification by chromatography.

b) N-(t-BuO₂C—CH₂)-N-Boc-(D)-Cha-Pro-NH—CH₂-4-(2-CSNH₂)-Thioph

N-(t-BuO₂C—CH₂)-N-Boc-(D)-Cha-Pro-NH—CH₂-4-(2-CN)-thioph (6.95 g, 11.53mmol) were [sic] dissolved in 40 ml of pyridine and 7 ml oftriethylamine, the solution was saturated with hydrogen sulfide at 0-5°C. (green solution) and left to stand at room temperature over theweekend. After concentration in vacuo at 35° C./35 mbar, the yellow oilyresidue was taken up in 200 ml of ether and washed four times with ineach case 20 ml of 20% strength sodium hydrogen sulfate solution, twicewith in each case 20 ml of saturated sodium hydrogen carbonate solutionand with 20 ml of water, and the organic phase was dried over soduimsulfate and concentrated in vacuo. 6.74 g resulted as a yellow solidfoam.

c) N-(t-BUO₂C—CH₂)-N-Boc-(D)-Cha-Pro-NH—CH₂-4-(2-C(SCH₃)NH)-ThiophHydroiodide

The crude product ofN-(t-BuO₂C—CH₂)-N-Boc-(D)-Cha-Pro-NH—CH₂-4-(2-CSNH₂)-thioph (6.74 g,10.58 mmol) was introduced into 65 ml of methylene chloride, 9.01 g (4.0ml, 63.5 mmol) of methyl iodide were added, and the mixture was left tostand overnight at room temperature. It was then concentrated in vacuounder mild conditions, 8.36 g resulting as a yellow solid foam.

d) N-(t-BuO₂C—CH₂)-N-Boc-(D)-Cha-Pro-NH—CH₂-4-(2-C(NH₂)NH)-ThiophHydroiodide

The crude product ofN-(t-BU02C—CH₂)-N-Boc-(D)-Cha-Pro-NH-CH₂-4-(2-C(SCH₃)NH)-thiophhydroiodide (8.36 g, max 10.58 mmol) together with 16.3 g (21.16 mmol)of a 10% strength ammonium acetate solution in methanol was stirredovernight at room temperature. Since the precursor had not reactedcompletely, another 1.63g of the 10% strength ammonium acetate solutionwere added and the mixture was again stirred overnight. Afterevaporation of the solvent in vacuo in a rotary evaporator, the residuewas taken up in methylene chloride, insoluble excess ammonium acetatewas removed by filtration, and the liquid was again concentrated invacuo, 7.12 g of the required product resulting as:a yellow solid foam.

e) HOOC—CH₂-(D)-Cha-Pro-NH—CH₂-4-(2-am)-Thioph Hydroacetate

The crude product ofN-(t-BuO₂C—CH₂)-Boc-(D)-Cha-Pro-NH—CH₂-4-(2-C(NH₂)NH)-thiophhydroiodide, which resulted from the above experiment, was dissolved in100 ml of methylene chloride, 24.5 ml of etheric hydrochloric acidsolution (approx. 5 N) were added, and the mixture was stirred overnightat room temperature. The resulting suspension was concentrated in vacuoand codistilled twice with methylene chloride, and the residue wasconverted into the acetate salt by means of acetate ion exchanger(Fluka, Product No. 00402), with 4.92 g being obtained. 2.5 g of thiswere purified by means of MPLC (RP-18, acetonitrile/water) and thefractions were freeze-dried. 1.23 g of the target product were obtainedas amorphous white solid.

FAB-MS (M+H⁺): 464.

EXAMPLE 3 N-(Hydroxycarbonylmethylene)-(D)-cyclohexylalanyl-pipecolicAcid [5-(2-Amidino)thienylmethyl]amide Hydroacetate

a) H-Pic-NH—CH₂-5-(2-CN)-Thioph

Boc-Pic-OH (10.1 g, 44.05 mmol) and 5-H₂N—CH₂-thioph-2-CN hydrochloride(8.54 g, 48.88 mmol) were dissolved in dichloromethane (150 ml), andethyl diisopropylamine (53.2 ml, 311.08 mmol) and a 50% strengthsolution of propanephosphonic anhydride in ethyl acetate (46 ml, 217mmol) were added at 0° C. After the reaction mixture had been stirredfor 1 hour at 0° C. and for 1 hour at room temperature, it was dilutedwith dichloromethane and washed with 20% strength sodium hydrogensulfate solution (4×), sodium hydrogen carbonate solution (3×) andsaturated sodium chloride solution (1×). After drying over sodiumsulfate and removal of the desiccant by filtration, the solvent wasdistilled out under water pump vacuum. To eliminate the Boc group, theresidue (18.41 g) was treated with 200 ml of isopropanol and 50 ml of6.8N isopropanolic hydrochloric acid solution and stirred overnight atroom temperature. The mixture was then evaporated to dryness andcodistilled twice with dichloromethane, and the residue was extracted bystirring with ether. 12.7 g of the required product resulted as a palebrown powder.

b) HOOC-H₂-(D)-Cha-Pic-NH—CH₂-5-(2-am)-Thioph Hydroacetate

This compound was prepared by coupling the two building blocksN-(t-BuO₂C—CH₂)-N-Boc-(D)-Cha-OH and H-Pic-NH—CH₂-5-(2-CN)-thiophanalogously to Example 2a). The reaction to give the end productHOOC—CH₂-(D)-Cha-Pic-NH—CH₂-5-(2-m)-thioph hydroacetate was carried outanalogously to Example 2b) to d), FAB-MS (M+H⁺): 478.

EXAMPLE 4N-(Hydroxycarbonylmethylene)-(D)-cyclohexylglycylprolyl-[2-(4-amidino)thienylmethyl]amideHydrochloride

a) HOOC—CH₂-(D)-Chg-Pyr-NH—CH₂-4-(2-am)-Thioph Dihydrochloride

This compound was prepared over several steps analogously to Examples 2and 3 starting from Boc-Pyr-OH, 4-H₂N—CH₂-thioph-2-N×HCl andN-(t-BuO₂-CH₂)-Boc-(D)-Chg-OH.

b) HOOC—CH₂-(D)-Chg-Pro-NH—CH₂-4-(2am)-Thioph Dihydrochloride

1.1 g (2.11 mmol) of HOOC—CH₂-(D)-Chg-Pyr-NH—CH₂-4-(2-am)-thiophdihydrochloride were dissolved in a mixture of 30 ml of water and 10 mlof glacial acetic acid, 0.5 g of 10% palladium on active charcoal wasadded, and the mixture was hydrogenated for 8 hours at room temperatureunder slightly elevated pressure. After the catalyst had been exchanged,hydrogenation continued for 8 hours, the catalyst was filtered off withsuction, the mixture was filtered through Celite® and theaqueous-organic phase was subsequently freeze-dried. 0.86 g of therequired product was obtained as a white amorphous solid.

FAB-MS (M+H⁺): 450.

As an alternative to the procedure described here, Boc-proline may beemployed directly instead of Boc-(L)-3,4-dehydroproline, which meansthat the hydrogenation step can be dispensed with.

EXAMPLE 5N-(Hydroxycarbonylmethylene)-(D)-cyclohexylglycylprolyl-[2-(4-amidino)thiazolylmethyl]amide

This compound can be prepared analogously to Example 1 starting fromN-(t-BuO₂C—CH₂)-N-Boc-(D)-Chg-Pro-OH and 2-H₂N—CH₂-thiaz-4-CSNH₂.

EXAMPLE 6N-(Hydroxycarbonylmethylene)-(D)-cyclohexylalanylprolyl-[4-(2-amidino)thienylmethyl]amide

This compound can be prepared analogously to Example 2 starting fromN-(t-BuO₂C—H₂)-N-Boc-(D)-Cha-Pro-OH and 4-H₂N—CH₂-thioph-2-CN oranalogously to Example 4 by hydrogenatingHOOC-CH₂-(D)-cha-Pyr-NH—CH₂-4-(2-am)-thioph.

EXAMPLE 7N-(Hydroxycarbonylmethylene)-(D)-cyclohexylalanylprolyl-[5-(2-amidino-3,4-dimethyl)thienylmethyl]amide

Starting from 5-H₂N—CH₂-(3,4-Me₂)-thioph-2-CONH₂ andN-(t-BuO₂C—CH₂)-N-Boc-(D)-Cha-Pro-OH, this compound can be convertedinto N-(t-BuO₂C—H₂)-N-Boc-(D)-Cha-Pro-NH—CH₂-5-(2-CONH₂-3,4-Me₂)-thiophanalogously to Example 2a). After dehydration of the amide withtrifluoroacetic anhydride and diisopropylethylamine in methylenechloride to give the nitrile functionality, the amidine functionalitycan be constructed analogously to Example 2 and the protective groupscan subsequently be eliminated.

EXAMPLE 8aN-(Hydroxycarbonylmethylene)-(D)-cycloheptylglycylprolyl-[4-(2-amidino)thienylmethyl]amideHydroacetate EXAMPLE 8bN-(Hydroxycarbonylmethylene)-(L)-cycloheptylglycylprolyl-[4-(2-amidino)thienylmethyl]amideHydroacetate

Starting from 4-H₂N—CH₂-thioph-2-CN, Boc-Pyr-OH andN-(t-BuO₂C—CH₂)-N-Boc-(D,L)-Cheg-OH, these compounds can be preparedanalogously to Example 3, hydrogenated in a final step analogously toExample 4 and subsequently separated by means of MPLC (RP 18,acetonitrile/water). If Boc-Pro-OH is employed in the synthesis insteadof Boc-Pyr-OH, the hydrogenation step can be dispensed with.

EXAMPLE 9aN-(Hydroxycarbonylmethylene)-(D)-cyclopentylglycylprolyl-[4-(2-amidino)thienylmethylamide Hydroacetate EXAMPLE 9bN-(Hydroxycarbbnylmethylene)-(L)-cyclopentylglycylprolyl-[4-(2-amidino)thienylmethyl]amideHydroacetate

Starting from 4-H₂N—CH₂-thioph-2-CN, Boc-Pyr-OH andN-(t-BuO₂C—CH₂)-N-Boc-(D,L)-Cpg-OH, these compounds can be preparedanalogously to Example 3, hydrogenated in a final step analogously toExample 4 and subsequently separated by means of MPLC (RP 18,acetonitrile/water). If Boc-Pro-OH is employed in the synthesis insteadof Boc-Pyr-OH, the hydrogenation step can be dispensed with.

EXAMPLE 10N-(Hydroxycarbonylmethylene)-(D)-cyclohexylalanylprolyl-[4-(2-amidino)thiazolylmethyl]amide

This compound can be prepared analogously to Example 1 starting 5 fromN-(t-BuO₂C—H₂)-Boc-(D)-Cha-Pro-OH and 4-H₂N—CH₂-thiaz-2-CSNH₂.

EXAMPLE 11N-(Hydroxycarbonylmethylene)-(D)-cyclohexylglycylprolyl-[4-(2-amidino)thiazolylmethyl]amide

This compound can be prepared analogously to Example 1 starting fromN-(t-BuO₂C—H₂)-N-Boc-(D)-Chg-Pro-OH and 4-H₂N—CH₂-thiaz-2-CSNH₂.

EXAMPLE 12 N-(Hydroxycarbonylmethylene)-(D)-cyclohexylalanylprolyl5-(3-Amidino)isoxazolylmethyl]amide

Starting from N-(t-BuO₂C—H₂)-N-Boc-(D)-Cha-Pro-OH and5-H₂N—CH₂-isox-3-CONH₂, this compound can be converted analogously toExample 2a) intoN-(t-BuO₂C—CH₂)-N-Boc-(D)-Cha-Pro-NH—CH₂-5-(3-CONH₂)-isox. Afterdehydration of the amide to give the nitrile functionality usingtrifluoroacetic anhydride and diisopropylethylamine in methylenechloride, the amidine functionalities can be constructed analogously toExample 20 by reaction with ammonia and acetylcysteine and theprotective groups can subsequently be eliminated.

EXAMPLE 13N-[5-(3-Amidino)thienylmethyl]-1-[N-(hydroxycarbonylmethylene)-(D)-cyclohexylglycyl]azetidine-2-caboxamide[sic]

a)N-[5-(3-Cyano)thienylmethyl]-1-[N-t-butoxycarbonyl)-(D)-cyclohexylglycyl]azetidine-2-carboxamide

5.9 g (45.8 mmol) of diisopropylethylamine and subsequently 11.5 ml(14.9 mmol) of 50% strength propanephosphonic anhydride-solution inethyl acetate were added dropwise at −5° C. to a solution of 3.9 g (11.5mmol) of1-[N-(t-butoxycarbonyl)-(D)-cyclohexylglycyl]azetidine-2-carboxylic acid(WO 9429336) and 2 g (11.5 mmol) of 5-aminomethyl-3-cyanothiophenehydrochloride in 40 ml of-methylene chloride. Stirring was continued for2 hours, during which process the temperature climbed to 10° C. Theorganic phase was washed with water, 5% strength sodium bicarbonatesolution and 5% strength citric acid solution, dried over sodium sulfateand evaporated to dryness. Purification of the residue by columnchromatography (silica gel, eluent: ethyl acetate) afforded 4.5 g (85%of theory) of white, amorphous powder, FAB-MS: 461 (M+H⁺).

b)N-[5-(3-Cyano)thienylmethyl]-1-[N-(t-butoxycarbonylmethylene)-(D)-cyclohexylglycyl]azetidine-2-carboxamide4.5 g (3.8 mmol) of the above compound were dissolved in 70 ml ofisopropanol, 12.3 ml of 4N hydrochloric acid in dioxane were added, andthe mixture was left to stand overnight at room temperature. After thesolvent had been distilled out, the residue was dissolved in methylenechloride and the mixture was extracted 3× with water. The combinedaqueous extracts were rendered alkaline with 1N sodium hydroxidesolution, the oily base [sic] which separated out was extracted 3× withmethylene chloride, and the solvent was subsequently distilled out. 2.9g (8 mmol) of oil remained. This was dissolved in 50 ml of methylenechloride and 10 ml of acetonitrile and, after addition of 2.1 g (16mmol) of diisopropylethylamine and 1.5 g (7.6 mmol) of t-butyl bromo-acetate, left to stand for 24 hours at room temperature.

The organic phase was washed in each case 2× with 5% strength citricacid solution, 5% strength sodium hydrogen carbonate solution and waterand dried over sodium sulfate and the solvent was distilled out. 3.3 g(92% of theory) of pale yellowish oil remained. FAB-MS: 475 (M+H⁺).

c)N-[5-(3-Amidino)thienylmethyl]-1-[N-(hydroxycarbonylmethylene)-(D)-cyclohexylglycyl]azetidine-2-carboxamide

The above product was coverted into the amidine analogously to Example2. The resulting crude amidine contained substantial amounts of asecondary product and had to be purified by column chromatography(silica gel, eluent: methylene chloride:methanol:acetic acid=24:6:1.5).0.9 g of white, amorphous powder was isolated. The Boc protective groupand the t-butyl ester group were eliminated by leaving the powder tostand for 12 hours in 3N hydrochloric acid. After finally removing thehydrochloric acid by distillation with addition of toluene, thehydrochloric residue was converted into the betaine by chromatographyover a silica gel column with a methanol/25% ammonia eluent (50/2.5).0.45 g of white, amorphous powder resulted, FAB-MS: 463 (M+H⁺).

EXAMPLE 14N-(Hydroxycarbonylmethylene)-(D)-yclohexylalanylprolyl-[5-(3-amidino)furylmethyl]amideHydroacetate

a) Prolyl-[5-(3-cyano)furylmethyl]amide Hydrochloride

3.05 g (14 mmol) of Boc-Pro-OH and 6.11 g (47.3 mmol) ofethyldiisopropylamine were added at room temperature to a suspension of2.5 g (15.8 mmol) of 5 minomethyl-3-cyanofuran hydrochloride in 50 ml ofdichloromethane. 15.8 ml (74.5 mmol) of a 50% strength solution ofpropanephosphonic anhydride in ethyl acetate were added dropwise at 5°C., with gentle cooling. After stirring for 30 minutes at roomtemperature, the mixture was diluted with ethyl acetate and washed threetimes with 5% strength citric acid solution, three times with saturatedsodium hydrogen carbonate solution and once with saturated sodiumchloride solution. After drying over sodium sulfate and removal of thedesiccant by filtration, the solvent was distilled out under water pumpvacuum. 4.3 g (86%) of a pale yellow oil resulted, and this was directlyreacted further.

The oil obtained in accordance with a) (4.3 g, 13.5 mmol) was dissolvedin 40 ml of ethyl acetate to eliminate the Boc group, and saturated withhydrogen chloride at 0° C. The reaction mixture was stirred overnightand evaporated to dryness in a rotary evaporator. 3.4 g (99%) of thetitle compound resulted.

b)N-(tert-Butoxycarbonylmethylene)-(N-Boc)-(D)-cyclohexylalanylprolyl-[5-(3-cyano)furylmethyl]amide

2.58 g (6.7 mmol) of t-BuO₂C—H₂-Boc-(D)-Cha-OH and 1.7 g (6.7 mmol) ofprolyl-[5-(3-cyano)furylmethyl]amide hydrochloride were suspended in 20ml of dichloromethane and 3.45 g (26.8 mmol) of ethyldiisopropylaminewere added. The reaction mixture was cooled to approx. 5° C., and 6.7 mlof a 50% strength solution of propanephosphonic anhydride in ethylacetate was added dropwise, during which process the solution turnedclear. After stirring overnight at room temperature, the mixture wasdiluted with ethyl acetate and then washed in each case three times with20% strength sodium hydrogen sulfate solution, saturated sodium hydrogencarbonate solution and once with saturated sodium chloride solution.After drying over sodium sulfate, the desiccant was removed and thesolvent was then distilled out under water pump vacuum. 3.7 g of therequired product resulted as an oil.

c)N-(tert-Butoxycarbonylmethylene)-(N-Boc)-(D)-cyclohexylalanylprolyl-[5-(3-midothiocarbonyl)furylmethyl]amide

The product obtained in accordance with b) was dissolved in pyridine (30ml) and triethylamine (15 ml). The reaction mixture was saturated withhydrogen sulfide at room temperature and stirred overnight at roomtemperature. The excess hydrogen sulfide was displaced by nitrogen andthe reaction mixture was poured into 300 ml of ice-cold 5% strengthsodium hydrogen sulfate solution. The mixture was extracted three timeswith ethyl acetate and the combined organic phases were washed once morewith 5% strength sodium hydrogen sulfate solution. After drying oversodium sulfate, the solvent was distilled out under water pump vacuum.The resulting crude product (3.3 g) was employed in the next stepwithout further purification.

d)N-(tert-Butoxycarbonylmethylene)-(N-Boc)-(D)-cyclohexylalanylprolyl-[5-(3-S-methyliminothiocarbonyl)furylmethyl]-amideHydroiodide

The crude product obtained in accordance with c) was dissolved in 50 mlof acetone, and 8.3 g (58.7 mmol) of methyl iodide were added. Afterstirring overnight at room temperature, the solvent was distilled outunder water pump vacuum. The residue was dissolved in a little ethylacetate and the solution was added dropwise to diisopropyl ether, duringwhich process a precipitate formed which was filtered off with suctionand washed with diisopropyl ether. Drying at room temperature in vacuoresulted in 3.3 g of a solid foam.

e)N-(tert-Butoxycarbonylmethylene)-(N-Boc)-(D)-cyclohexylalanylprolyl-[5-(3-amidino)furylmethyl]amideHydroacetate

The crude product obtained in accordance with d) (3.3 g, 4.3 mmol) wasdissolved in 40 ml of acetonitrile, 0.99 g (12.9 mmol) of ammoniumacetate were added, and the mixture was stirred for two hours at 40° C.The solvent was then distilled out under water pump vacuum, the residuewas taken up in diethyl ether, the salts were filtered off with suctionand the filtrate was concentrated. The crude product was purified bymeans of reversed phase HPLC (acetonitrile/water and acetic acidbuffer), which resulted in 991 mg of a yellow solid foam.

f)N-(Hydroxycarbonylmethylene)-(D)-cyclohexylalanylprolyl-[5-(3-amidino)furylmethyl]amideHydroacetate

20 ml of a 1N aqueous hydrochloric acid solution were added to theproduct obtained in accordance with e) (991 mg, 1.68 mmol). Afterstirring for three hours at 45° C., the mixture was diluted with waterand the resulting mixture was freeze-dried. The resulting crude productwas dissolved in methanol and converted into the acetate salt over anion exchanger (Fluka, Product No. 00402). 484 mg of the required productresulted.

FAB-MS (M+H⁺): 446.

EXAMPLE 15N-(Hydroxycarbonylmethylene)-(D)-cyclohexylglycylprolyl-(5-(3-amidino)furylmethyl]amideHydroacetate

FAB-MS (M+H⁺): 434.

This can be prepared analogously to Example 14,N-(tert-butoxycarbonylmethylene)-(N-Boc)-(D)-cyclohexylglycine beingemployed instead ofN-(tert-butoxycarbonylmethylene)-(N-Boc)-(D)-cyclohexylalanine in b).

EXAMPLE 16N-(Hydroxycarbonylmethylene)-(D)-cyclohexylalanylprolyl-[5-(2-amidino-1-methyl)pyrrolylmethyl]amide

a) N-Boc-N-(tert-Butyloxycarbonylmethylene)-(D)-cyclohexylalanylproline(2.5 g, 5.18 mmol) was dissolved in dry methylene chloride (30 ml),cooled to −10° C., and N-ethyldiisopropylamine (3.9 ml, 22.27 mmol) wasadded at this temperature. After stirring for 5 minutes, a solution of5-aminomethyl-1-methylpyrrole-2-carbonitrile (0.7 g, 5.18 mmol) inmethylene chloride (15 ml) was added. A 50% strength propanephosphonicanhydride solution in ethyl acetate (4.6 ml, 6.21 mmol) was subsequentlyadded dropwise in the course of 20 minutes. After stirring for 90minutes at −10° C. to 0° C., the mixture was diluted with methylenechloride and washed 2× with saturated sodium hydrogen carbonate solution(in each case 15 ml), 2× with 5% strength citric acid solution (in eachcase 15 ml) and 1× with saturated sodium chloride solution (15 ml).After drying over sodium sulfate, the mixture was concentrated in vacuoand the crude product was purified by chromatography (silica gel,methylene chloride:methanol=95:5). Yield: 2.3 g (74%).

b) The product obtained in accordance with a) (2.3 g, 3.83 mmol) wasdissolved in a mixture of dry methylene chloride and methanol (1:1, 50ml), hydroxylamine hydrochloride (664 mg, 9.56 mmol) andN-ethyldiisopropylamine (4 ml, 23.0 mmol) were added, and the mixturewas stirred for 7 hours at 40° C. and subsequently for 48 hours at roomtemperature. The solvent was distilled out in vacuo, water was added tothe residue, and the mixture was acidified to pH 5 with acetic acid. Theaqueous solution was extracted with methylene chloride (2×) and ethylacetate (1×). The combined organic phases were dried over sodium sulfateand the solvent was distilled out in vacuo. The crude product waspurified by chromatography (silica gel, methylenechloride:methanol=95:5). Yield: 1.6 g (white foam, 66%),

FAB-MS (M+H⁺): 633.

c) The product obtained in accordance with b) (1.6 g, 2.53 mmol) wasdissolved in dry methanol (35 ml), acetic acid (0.3 ml, 5.06 mmol) andRaney nickel (84 mg) were added, and the mixture was hydrogenated at 50°C. under 1 atmosphere of hydrogen (2.5 hours). After cooling, thecatalyst was removed by filtration through Celite® and the filtrate wasconcentrated in vacuo. Yield: 1.7 g (white foam, 99%), FAB-MS (M+H⁺):617.

d) The product obtained in accordance with c) (1.7 g, 2.50 mmol) wasdissolved in dry methylene chloride (50 ml). The solution was cooled to0° C. and saturated with dry HCl gas. After stirring for 2 hours, thesolvent was distilled out in vacuo and the crude product was purified bychromatography (RP18, acetonitrile:water=1:9 with addition of 0.1%acetic acid).

Yield: 760 mg (57%), melting point: 184-185° C., FAB-MS (M+H⁺): 461.

EXAMPLE 17N-(Hydroxycarbonylmethylene)-(D)-cyclohexylalanylprolyl-[2-(4-amidino-1-methyl)pyrrolemethyl]amidewas prepared analogously to Example 16, FAB-MS (M+H⁺): 461. EXAMPLE 18N-(Hydroxycarbonylmethylene)-(D)-cyclohexylalanylprolyl-[4-(2-amidino-1-methyl)pyrrolylmethyl]amidecan be Prepared Analogously to Example 16. EXAMPLE 19N-(tert-Butoxycarbonylmethylene)-(N-Boc)-(D)-cyclohexylalanylprolyl-[2-(4-amido)oxazolylmethyl]amideHydrochloride can be Prepared Analogously to Example 1 Starting FromN-(t-BuO₂C—CH₂-N-Boc-(D)-Cha-Pro-OH and2-Aminomethyl-4-thiocarboxamidoxazole EXAMPLE 20N-(Hydroxycarbonylmethylene)-(D)-cyclohexylalanylprolyl-[5-(3-amidino-1-methyl)pyrazolylmethyl]amideHydrochloride

a)N-(tert-Butoxycarbonylmethylene)-(N-Boc)-(D)-cyclohexylalanylprolyl-[5-(3-amido-1-methyl)pyrazolylmethyl]amide

N-(t-BuO₂C—CH₂)-N-Boc-(D)-Cha-Pro-OH (1.25 g, 2.59 mmol) was introducedinto dichloromethane (30 ml). Diisopropylethylamine (1.95 ml, 11.16mmol) was added dropwise at −10° C. A solution of1-methyl-5-aminomethylpyrazole-3-carboxamide (0.4 g, 2.59 mmol) intetrahydrofuran (20 ml) was subsequently added. After stirring for 5minutes, a 50% strength solution of propanephosphonic anhydride ethylacetate solution [sic] (2.36 ml, 3.11 mmol), as well as dichloromethane(5 ml), were added dropwise in the course of 5 minutes. After stirringfor 45 minutes at 0° C., the mixture was warmed at RT for 12 hours. Thesolvent was removed in a rotary evaporator, the residue was taken up indichloromethane and the mixture was washed 2× with saturated sodiumhydrogen carbonate solution, 2× with 5% strength citric acid solutionand 1× with saturated sodium chloride solution. After drying over sodiumsulfate, the solvent was removed in a rotary evaporator. The crudeproduct was purified by chromatography (RP-18, acetonitrile, water).

Yield: 220 mg (14%). FAB-MS (M+H⁺): 619.

b)N-(tert-Butoxycarbonylmethylene)-(N-Boc)-(D)-cyclohexylalanylprolyl-[5-(3-cyano-1-methyl)pyrazolylmethyl]amide

The product obtained in accordance with a) (220 mg, 0.36 mmol) wasdissolved in dichloromethane (15 ml), and diisopropylethylamine (0.17ml, 0.96 mmol) was added at −10° C. After stirring for 5 minutes, asolution of trifluoroacetic anhydride (0.057 ml, 0.41 mmol) indichloromethane (1 ml) was added dropwise. After 1 hour at 0° C., themixture was diluted with dichloromethane and washed 2× with saturatedsodium hydrogen carbonate solution, 2× with 5% strength citric acidsolution and 1× with saturated sodium chloride solution. After dryingover sodium sulfate, the solvent was removed in a rotary evaporator.Yield: 180 mg (84%).

c)N-(tert-Butoxycarbonylmethylene)-(N-Boc)-(D)-cyclohexylalanyprolyl-[5-(3-amidino-1-methyl)pyrazolylmethyl]amideHydroacetate

The product obtained in accordance with b) (180 mg, 0.3 mmol) wasdissolved in methanol (1 ml), and acetylcysteine (52.8 mg, 0.32 mmol)was added. Ammonia was subsequently passed in at 35° C. until thereaction was complete. The solvent was removed in a rotary evaporatorand the crude product was converted into the acetate using an ionexchanger (acetate on polymeric support, Fluka 00402). The crude productwas purified by chromatography (RP-18, acetonitrile, water). Yield: 50mg (16%), FAB-MS (M+H⁺): 618.

d)N-(Hydroxycarbonylmethylene)-(D)-cyclohexylalanylprolyl-[5-(3-amidino-1-methyl)pyrazolylmethyl]amideHydrochloride

The product obtained in accordance with c) (50 mg, 0.081 mmol) wasdissolved in dichloromethane (5 ml), and 5M hydrochloric acid in diethylether (0.147 ml) was added. After stirring for 12 hours at RT, thesolvent was removed in a rotary evaporator, and the product was taken upin water and lyophilized. Yield: 40 mg (92%), FAB-MS (M+H⁺): 462.

EXAMPLE 21 N-(Hydroxycarbonylmethylene)-(D)-cyclohexylalanylprolyl-[5-(3midino)-1,2,4-oxadiazolylmethyl amide Hydrochloride can be PreparedAnalogously to Example 20 Starting From t-BuO₂C—CH₂-(Boc)-(D)-Cha-Pro-OHand 5-Amino-3-cyano-1,2,4-oxadiazole EXAMPLE 22N-(Hydroxycarbonylmethylene)-(D)-cyclohexylglycylprolyl-[5-(2-amidino-3-methyl)thienylmethyl]amide

This compound was synthesized starting from N-(t-BuO₂C—CH₂)N-Boc-(D)-Chg-Pro-OH and 5-aminomethyl-3-methylthiophene-2-carbonitrilehydrochloride by a method similar to that described in Ex. 2a)-e).

FAB-MS (M+H⁺): 464.

EXAMPLE 23N-(Hydroxycarbonylmethylene)-(D)-cyclohexylalanylprolyl-[5-(2-amidino-3-methyl)thienylmethyl]amide

This compound was synthesized starting fromN-(t-BuO₂C—CH₂)-N-Boc-(D)-Cha-Pro-OH and5-aminomethyl-3-methylthiophene-2-carbonitrile hydrochloride by a methodsimilar to that described in Ex. 2 a)-e).

FAB-MS (M+H⁺): 478.

EXAMPLE 24 N-(Hydroxycarbonylmethylene)-(D)-cyclohexylalanylprolyl-[5-(3amidino-1-methyl)triazylmethyl]amide

This compound was synthesized starting fromN-(t-Bub₂C—CH₂)-N-Boc-(D)-Cha-Pyr-OH and5-aminomethyl-1-methyl-1H-[1,2,4]triazole-3-carboxamide. First, a methodsimilar to that described in Ex. 20a)-d) was followed, andN-(hydroxycarbonylmethylene)-(D)-cyclohexylalanyldehydroprolyl-[5-(3-amidino-1-methyl)triazylmethyl]amidewas obtained. This compound was converted into the title compound by amethod similar to that described in Ex. 4b).

FAB-MS (M+H⁺): 463.

EXAMPLE 25 N-(Hydroxycarbonylmethylene)-(D)-cyclohexylglycylprolyl-[5-(3amidino-1-methyl)triazylmethyl]amide

This compound was synthesized starting fromN-(t-BuO₂C—CH₂)-N-Boc-(D)-Chg-Pyr-OH and5-aminomethyl-1-methyl-1H-[1,2,4]triazole-3-carboxamide. First, a methodsimilar to that described in Ex. 20a)-d) was followed, andN-(hydroxycarbonylmethylene)-(D)-cyclohexylglycyldehydroprolyl-[5-(3-amidino-1-methyl)triazylmethyl]amidewas obtained. This compound was converted into the title compound by amethod similar to that described in Ex. 4b).

FAB-MS (M+H⁺): 449.

EXAMPLE 26N-(Hydroxycarbonylmethylene)-(D)-cyclohexylalanylprolyl-[5-(3-amidino-4-chloro)thienylmethyl]amide

This compound was synthesized starting fromN-(t-BuO₂C—CH₂)-N-Boc-(D)-Cha-Pro-OH and5-aminomethyl-4-chlorothiophene-3-thiocarboxamide hydrochloride by amethod similar to that described in Example 1a)-d).

ESI-MS (M+H⁺): 498.

EXAMPLE 27N-(Ethoxycarbonylmethylene)-(D)-cyclohexylalanylprolyl-[5-(3-amidino-4-chloro)thienylmethyl]amide

This compound was synthesized by passing hydrogen chloride to saturationinto a solution, cooled to 0° C., of 100 mg (0.186 mmol) ofN-(hydroxycarbonylmethylene)-(D)-cyclohexylalanylprolyl-[5-(3amidino-4-methyl)thienyl]methylamide(Example 26 above) in 10 ml of ethanol and stirring for five hours atroom temperature. The mixture was concentrated and co-distilled threetimes with a little toluene each time in order to remove residualhydrogen chloride. The residue (89 mg, 85%) was dissolved in ethanol andconverted into the corresponding acetate by means of an acetate ionexchanger (Fluka, Product No. 00402). FAB-MS (M+H⁺): 506.

EXAMPLE 28N-(Hydroxycarbonylmethylene)-(D)-cyclohexylalanylproly-1[5-(3-amidino-4-methyl)thienylmethyl]amide

This compound was synthesized starting fromN-(t-BuO₂C—CH₂)-N-Boc-(D)-Cha-Pro-OH and5-aminomethyl-4-methylthiophene-3-thiocarboxamide hydrochloride by amethod similar to that described in Example 1a)-d). ESI-MS (M+H⁺): 478.

EXAMPLE 29N-(Ethoxycarbonylmethylene)-(D)-cyclohexylalanylprolyl-[5-(3-amidino-4-methyl)thienylmethyl]amide

This compound was synthesized by passing hydrogen chloride to saturationinto a solution, cooled to 0° C., of 100 mg (0.186 mmol) ofN-(hydroxycarbonylmethylene)-(D)-cyclohexylalanylprolyl-[5-(3-amidino-4-methyl)thienyl]methylamide(Example 28 above) in 10 ml of ethanol and stirring the mixture for 5hours at room temperature. The mixture was concentrated and co-distilledthree times with a little toluene each time in order to remove residualhydrogen chloride. The residue (89 mg, 85%) was dissolved in ethanol andconverted into the corresponding acetate by means of an acetate ionexchanger (Fluka, Product No. 00402).

FAB-MS (M+H⁺): 506.

EXAMPLE 30N-(Hydroxycarbonylmethylene)-(D)-cyclohexylalanylprolyl-[5-(2-amidino-3-chloro)thienylmethyl]amide

This compound can be prepared by the following reaction sequence:coupling of N-(t-BuO₂C—CH₂)-Boc-(D)-Cha-Pro-OH with5-H₂N—CH₂-(2-CN-3-Cl)-thioph to giveN-(t-BuO₂C—CH₂)-Boc-(D)-Cha-Pro-NH—CH₂-5-(2-CN-3-Cl)-thioph, amidineformation and subsequent elimination of the protective groups by amethod similar to that described in Example 2.

EXAMPLE 31N-(Hydroxycarbonylmethylene)-(D)-yclohexylglycylprolyl-[5-(2-amidino-3-chloro)thienylmethyl]amide

This compound can be prepared by the following reaction sequence:coupling of N-(t-BuO₂C—CH₂)-N-BQc-(D)-Chg-Pro-OH with5-H₂N—CH₂-(2-CN-3-Cl)-thioph to giveN-(t-BuO₂C—CH₂)-N-Boc-(D)-Chg-Pro-NH—CH₂-5-(2-CN-3-Cl)-thioph, amidineformation and subsequent elimination of the protective groups by amethod similar to that described in Example 2.

EXAMPLE 32N-(Methoxycarbonylmethylene)-(D)-cyclohexylalanylprolyl-[4-(2-amidino)thienylmethyl]amide

This compound can be synthesized fromN-(t-BuO₂C—CH₂)-N-Boc-(D)-Cha-Pro-NH—CH₂-4-(2-am)-thioph by eliminationof the protective groups and transesterification (HCl in methanol atroom temperature).

EXAMPLE 33N-(Methoxycarbonylmethylene)-(D)-cyclohexylglycylprolyl[4-(2-amidino)thienylmethyl]amide

This compound can be synthesized fromN-(t-BuO₂C—CH₂)-N-Boc-(D)-Chg-Pro-NH—CH₂-4-(2-am)-thioph by eliminationof the protective groups and transesterification (HCl in methanol atroom temperature).

EXAMPLE 34N-(Methoxycarbonylmethylene)-(D)-cyclohexylalanyl-azetidine-2-carboxylicAcid 4-(2-amidino)thienylmethylamide

This compound can be synthesized fromN-(t-BuO₂C—CH₂)-N-Boc-(D)-Cha-Aze-NH—CH₂-4-(2-am)-thioph by eliminationof the protective groups and transesterification (HCl in methanol atroom temperature).

EXAMPLE 35N-(Methoxycarbonylmethylene)-(D)-cyclohexylglycyl-azetidine-2-carboxylicacid 2-(4-amidino)thienylmethylamide

This compound can be synthesized fromN-(t-BuO₂C—CH₂)-N-Boc-(D)-Chg-Aze-NH—CH₂-4-(2-am)-thioph by eliminationof the protective groups and transesterification (HCl in methanol atroom temperature).

EXAMPLE 36N-(Methoxycarbonylmethylene)-(D)-cyclohexylalanylprolyl-[2-(4-amidino)thiazolylmethylamide

This compound can be synthesized fromN-(t-BuO₂C—CH₂)-N-Boc-(D)-Cha-Pro-NH—CH₂-2-(4-am)-thiaz by eliminationof the protective groups and transesterification (HCl in methanol atroom temperature).

EXAMPLE 37N-(Methoxycarbonylmethylene)-(D)-cyclohexylglycylprolyl-[2-(4-amidino)thiazolylmethylamide

This compound can be synthesized fromN-(t-BuO₂C—CH₂)-N-Boc-(D)-Chg-Pro-NH—CH₂-2-(4-am)-thiaz by eliminationof the protective groups and transesterification (HCl in methanol atroom temperature).

EXAMPLE 38N-(Methoxycarbonylmethylene)-(D)-cyclohexylalanyl-azetidine-2-carboxylicacid 2-(4-amidino)thiazolylmethylamide

This compound can be synthesized fromN-(t-BuO₂C—CH₂)-N-Boc-(D)-Cha-Aze-NH—CH₂-2-(4-am)-thiaz by eliminationof the protective groups and transesterification (HCl in methanol atroom temperature).

EXAMPLE 39N-(Methoxycarbonylmethylene)-(D)-cyclohexylglycyl-azetidine-2-carboxylicacid 2-(4-amidino)thiazolylmethylamide

This compound can be synthesized fromN-(t-BuO₂C—CH₂)-N-Boc-(D)-Chg-Aze-NH—CH₂-2-(4-am)-thiaz by eliminationof the protective groups and transesterification (HCl in methanol atroom temperature).

EXAMPLE 40N-(Hydroxycarbonylmethylene)-(D)-cyclohexylalanylprolyl-[4-(2-hydroxyamidino)thienylmethyl]amide

This compound can be synthesized by reacting(t-BuO₂C—CH₂-)-(Boc)-(D)-Cha-Pro-NH—CH₂-(2-CN)-4-thioph withhydroxylamine hydrochloride (methanol, diisopropylethylamine, roomtemperature) and subsequent elimination of the protective groups (HCl indichloromethane at room temperature).

EXAMPLE 41N-(Methoxycarbonylmethylene)-(D)-cyclohexylalanylprolyl-[4-(2-hydroxyamidino)thienylmethyl]amide

This compound can be synthesized by reacting(t-BuO₂C—CH₂-)-(Boc)-(D)-Cha-Pro-NH—CH₂-(2-CN)-4-thioph withhydroxylamine hydrochloride (methanol, diisopropylethylamine, roomtemperature) and subsequent elimination of the protective groups andtransesterification (HCl in methanol at room temperature).

EXAMPLE 42N-(Ethoxycarbonylmethylene)-(D)-cyclbhexylalanylprolyl-[4-(2-hydroxyamidino)thienylmethyl]amide

This compound can be synthesized by reacting(t-BuO₂C—CH₂-)-(Boc)-(D)-Cha-Pro-NH—CH₂-(2-CN)-4-thioph withhydroxylamine hydrochloride (methanol, diisopropylethylamine, roomtemperature) and subsequent elimination of the protective groups andtransesterification (HCl in ethanol at room temperature).

EXAMPLE 43N-(Hydroxycarbonylmethylene)-(D)-cyclohexylglycylprolyl-[4-(2-hydroxyamidino)thienylmethyl]amide

This compound can be synthesized by reacting(t-BuO₂C—CH₂-)-(Boc)-(D)-Chg-Pro-NH—CH₂-(2-CN)-4-thioph withhydroxylamine hydrochloride (methanol, diisopropylethylamine, roomtemperature) and subsequent elimination of the protective groups (HCl indichloromethane at room temperature).

EXAMPLE 44N-(Methoxycarbonylmethylene)-(D)-cyclohexylglycylprolyl-[4-(2-hydroxyamidino)thienylmethyl]amide

This compound can be synthesized by reacting(t-BuO₂C—CH₂-)-(Boc)-(D)-Chg-Pro-NH—CH₂-(2-CN)-4-thioph withhydroxylamine hydrochloride (methanol, diisopropylethylamine, roomtemperature) and subsequent elimination of the protective groups andtransesterification (HCl in methanol at room temperature).

EXAMPLE 45N-(Ethoxycarbonylmethylene)-(D)-cyclohexylglycylprolyl-[4-(2-hydroxyamidino)thienylmethyl]amide

This compound can be synthesized by reacting(t-BuO₂C—CH₂-)-(Boc)-(D)-Chg-Pro-NH—CH₂-(2-CN)-4-thioph withhydroxylamine hydrochloride (methanol, diisopropylethylamine, roomtemperature) and subsequent elimination of the protective groups andtransesterification (HCl in ethanol at room temperature).

EXAMPLE 46N-(Hydroxycarbonylmethylene)-(D)-cyclohexylglycyl-azetidine-2-carboxylicAcid 4-(2-Hydroxyamidino)-thienylmethyl]amide

This compound can be synthesized by reacting(t-BuO₂C—CH₂-)-(Boc)-(D)-Chg-Aze-NH—CH₂-(2-CN)-4-thioph withhydroxylamine hydrochloride (methanol, diisopropylethylamine, roomtemperature) and subsequent elimination of the protective groups (HCl indichloromethane at room temperature).

EXAMPLE 47N-(Methoxycarbonylmethylene)-(D)-cyclohexylglycyl-azetidine-2-carboxylicAcid [4-(2-Hydroxyamidino)-thienylmethyl]amide

This compound can be synthesized by reacting(t-BuO₂C—CH₂-)-(Boc)-(D)-Chg-Aze-NH—CH₂-(2-CN)-4-thioph withhydroxylamine hydrochloride (methanol, diisopropylethylamine, roomtemperature) and subsequent elimination of the protective groups andtransesterification (HCl in methanol at room temperature).

EXAMPLE 48N-(Ethoxycarbonylmethylene)-(D)-cyclohexylglycyl-azetidine-2-carboxylicAcid [4-(2-Hydroxyamidino)-thienylmethyl]amide

This compound can be synthesized by reacting(t-BuO₂C—CH₂-)-(Boc)-(D)-Chg-Aze-NH—CH₂-(2-CN)-4-thioph withhydroxylamine hydrochloride (methanol, diisopropylethylamine, roomtemperature) and subsequent elimination of the protective groups andtransesterification (HCl in ethanol at room temperature).

EXAMPLE 49N-(Hydroxycarbonylmethylene)-(D)-cyclohexylalanyl-azetidine-2-carboxylicAcid [4-(2-Hydroxyamidino)-thienylmethyl]amide

This compound can be synthesized by reacting(t-BuO₂C—CH₂-)-(Boc)-(D)-Cha-Aze-NH—CH₂-(2-CN)-4-thioph withhydroxylamine hydrochloride (methanol, diisopropylethylamine, roomtemperature) and subsequent elimination of the protective groups (HCl indichloromethane at room temperature).

EXAMPLE 50N-(Methoxycarbonylmethylene)-(D)-cyclohexylalanyl-azetidine-2-carboxylicAcid 14-(2-Hydroxyamidino)-thienylmethyl]amide

This compound can be synthesized by reacting(t-BuO₂C—CH₂-)-(Boc)-(D)-Cha-Aze-NH—CH₂-(2-CN)-4-thioph withhydroxylamine hydrochloride (methanol, diisopropylethylamine, roomtemperature) and subsequent elimination of the protective groups andtransesterification (HCl in methanol at room temperature).

EXAMPLE 51N-(Ethoxycarbonylmethylene)-(D)-cyclohexylalanyl-azetidine-2-carboxylicAcid [4-(2-Hydroxyamidino)-thienylmethyl]amide

This compound can be synthesized by reacting(t-BUO₂C—CH₂-)-(Boc)-(D)-Cha-Aze-NH—CH₂-(2-CN)-4-thioph withhydroxylamine hydrochloride (methanol, diisopropylethylamine, roomtemperature) and subsequent elimination of the protective groups andtransesterification (HCl in ethanol at room temperature).

EXAMPLE 52N-(Hydroxycarbonylmethylene)-(D)-cyclohexylalanylprolyl-[2-(4-hydroxyamidino)thiazolylmethyl]amide

This compound can be synthesized by reacting(t-BUO₂C—CH₂-)-(Boc)-(D)-Cha-Pro-NH—CH₂-(4-CN)-2-thiaz withhydroxylamine hydrochloride (methanol, diisopropylethylamine, roomtemperature) and subsequent elimination of the protective groups (HCl indichloromethane at room temperature).

EXAMPLE 53N-(Methoxycarbonylmethylene)-(D)-cyclohexylalanylprolyl-[2-(4-hydroxyamidino)thiazolylmethyl]amide

This compound can be synthesized by reacting(t-BuO₂C—CH₂-)-(Boc)-(D)-Cha-Pro-NH—CH₂-(4-CN)-2-thiaz withhydroxylamine hydrochloride (methanol, diisopropylethylamine, roomtemperature) and subsequent elimination of the protective groups andtransesterirfication (HCl in methanol at room temperature).

EXAMPLE 54N-(Ethoxycarbonylmethylene)-(D)-cyclohexylalanylprolyl-(2-(4-hydroxyamidino)thiazolylmethyl]amide

This compound can be synthesized by reacting(t-BuO₂C—CH₂-)-(Boc)-(D)-Cha-Pro-NH—CH₂-(4-CN)-2-thiaz withhydroxylamine hydrochloride (methanol, diisopropylethylamine, roomtemperature) and subsequent elimination of the protective groups andtransesterification (HCl in ethanol at room temperature).

EXAMPLE 55N-(Hydroxycarbonylmethylene)-(D)-cyclohexylglycylprolyl-[2-(4-hydroxyamidino)thiazolylmethyl]amide

This compound can be synthesized by reacting(t-BuO₂C—CH₂-)-(Boc)-(D)-Chg-Pro-NH—CH₂-(4-CN)-2-thiaz withhydroxylamine hydrochloride (methanol, diisopropylethylamine, roomtemperature) and subsequent elimination of the protective groups (HCl indichloromethane at room temperature).

EXAMPLE 56N-(Methoxycarbonylmethylene)-(D)-cyclohexylglycylprolyl-[2-(4-hydroxyamidino)thiazolylmethyl]amide

This compound can be synthesized by reacting(t-BuO₂C—CH₂-)-(Boc)-(D)-chg-Pro-NH—CH₂-(4-CN)-2-thiaz withhydroxylamine hydrochloride (methanol, dilsopropylethylamine, roomtemperature) and subsequent elimination of the protective groups andtransesterification (HCl in methanol at room temperature).

EXAMPLE 57N-(Ethoxycarbonylmethylene)-(D)-cyclohexylglycylprolyl-[2-(4-hydroxyamidino)thiazolylmethyl]amide

This compound can be synthesized by reacting (t-BUO₂C—CH₂-)-5(Boc)-(D)-Chg-Pro-NH—CH₂-(4-CN)-2-thiaz with hydroxylamine hydrochloride(methanol, diisopropylethylamine, room temperature) and subsequentelimination of the protective groups and transesterification (HCl inethanol at room temperature).

EXAMPLE 58N-(Hydroxycarbonylmethylene)-(D)-cyclohexylglycyl-azetidine-2-carboxylicAcid [2-(4-Hydroxyamidino)-thiazolylmethyl]amide

This compound can be synthesized by reacting(t-BuO₂C—CH₂-)-(Boc)-(D)-Chg-Aze-NH—CH₂-(4-CN)-2-thiaz withhydroxylamine hydrochloride (methanol, diisopropylethylamine, roomtemperature) and subsequent elimination of the protective groups (HCl indichloromethane at room temperature).

EXAMPLE 59N-(Methoxycarbonylmethylene)-(D)-cyclohexylglycyl-azetidine-2-carboxylicAcid [2-(4-Hydroxyamidino)-thiazolylmethyl]amide

This compound can be synthesized by reacting(t-BuO₂C—CH₂-)-(Boc)-(D)-Chg-Aze-NH—CH₂-(4-CN)-2-thiaz withhydroxylamine hydrochloride (methanol, diisopropylethylamine, roomtemperature) and subsequent elimination of the protective groups andtransesterification (HCl in methanol at room temperature).

EXAMPLE 60N-(Ethoxycarbonylmethylene)-(D)-cyclohexylglycyl-azetidine-2-carboxylicAcid [2-(4-Hydroxyamidino)-thiazolylmethyl]amide

This compound can be synthesized by reacting(t-BuO₂C—CH₂-)-(Boc)-(D)-Chg-Aze-NH—CH₂-(4-CN)-2-thiaz withhydroxylamine hydrochloride (methanol, diisopropylethylamine, roomtemperature) and subsequent elimination of the protective groups (HCl inethanol at room temperature).

EXAMPLE 61N-(Hydroxycarbonylmethylene)-(D)-cyclohexylalanyl-azetidine-2-carboxylicAcid [2-(4-Hydroxyamidino)-thiazolylmethyl]amide

This compound can be synthesized by reacting(t-BuO₂C—CH₂-)-(Boc)-(D)-Cha-Aze-NH—CH₂-(4-CN)-2-thiaz withhydroxylamine hydrochloride (methanol, dlisopropylethylamine, roomtemperature) and subsequent elimination of the protective groups (HCl indichloromethane at room temperature).

EXAMPLE 62N-(Methoxycarbonylmethylene)-(D)-cyclohexylalanyl-azetidine-2-carboxylicAcid [2-(4-Hydroxyamidino)-thiazolylmethyl]amide

This compound can be synthesized by reacting(t-BuO₂C—CH₂-)-(Boc)-(D)-Cha-Aze-NH—CH₂-(4-CN)-2-thiaz withhydroxylamine hydrochloride (methanol, diisopropylethylamine, roomtemperature) and subsequent elimination of the protective groups andtransesterification (HCl in methanol at room temperature).

EXAMPLE 63N-(Ethoxycarbonylmethylene)-(D)-cyclohexylalanyl-azetidine-2-carboxylicAcid [2-(4-Hydroxyamidino)-thiazolylmethyl]amide

This compound can be synthesized by reacting(t-BuO₂C—CH₂-)-(Boc)-(D)-Cha-Aze-NH—CH₂-(4-CN)-2-thiaz withhydroxylamine hydrochloride (methanol, diisopropylethylamine, roomtemperature) and subsequent elimination of the protective groups andtransesterification (HCl in ethanol at room temperature).

EXAMPLE 64N-(Hydroxycarbonylmethylene)-(R)-cyclohexylalanyl-(3S)-2,3,4,5-tetrahydropyridazine-3-carboxylicAcid [4-(2-Amidino)thienylmethyl]amide Hydrochloride

a)N-Boc-N-(tert-Butoxycarbonylmethylene)-(D)-cyclohexylalanyl-(3S)-2,3,4,5-tetrahydropyridazine-3-carboxylicAcid

3.3 g (25.8 mmol) of diisopropylethylamine and 1.5 g (12.3 mmol) ofdimethylaminopyridine were added at −5° C. to a solution of 9.95 g (25.8mmol) of N-Boc-N-(tert-butoxy-carbonylmethylene)-(D)-cyclohexylalanine,7.4 g (25.8 mmol) of(S)-4-benzyl-3-[(S)-2,3,4,5-tetrahydro-3-pyridazinyl]-2-oxazolidinone(Y. Nakamura, C. Shin, Chem. Lett. 1991, 1953) and 7.4 g (38.7 mmol) ofEDC hydrochloride in 80 ml of CH₂Cl₂ and the mixture was stirred for 2hours at −5° C. and for 12 hours at room temperature.

The reaction solution was diluted with 200 ml of ether, washed with 5%strength citric acid solution, 5% strength NaHCO₃ solution and water,and, after the mixture had been dried and the solvent stripped off, theresidue was purified by column chromatography (methylene chlorideacetone [sic], 50/2.5). This gave 4.0 g (24% of theory) of a yellowishoil, FAB-MS (M+H⁺): 655. This was dissolved in 80 ml of THF and 27 ml ofwater, and, at 0° C., 2.8 ml of 30% strength H₂O₂ and 12.6 ml of 1N NaOHwere successively added dropwise and stirring was continued for 2.5hours. After addition of 15 g of a saturated aqueous Na₂S₂O₃ solution,the mixture was extracted with ether and the alkaline phase wasseparated off and acidified with 1M KHSO₄ solution. After repeatedextraction with ether, drying and removal of the solvent bydistillation, 2.2 g of a white amorphous powder remained.

b) HOOC—CH₂-(D)-Cha-(3S)-2,3,4,5-Tetrahydropyridazine-3-carboxylic Acid[2-(4am)Thienylmethyl]amide Hydrochloride

0.7 g (1.4 mmol) of the above acid and 0.3 g of2-aminomethyl-4-amidinothiophene dihydrochloride were suspended in 4 mlof DMF. After addition of 0.145 g (1.44 mmol) of N-methylmorpholine at0° C., almost complete solution took place, and 0.475 g (1.45 mmol) of0-[cyanoethoxycarbonylmethylene)amino]-N,N,N′,N′-tetramethyluroniumtetrafluoroborate[sic] (TOTU) and a further 0.14 g of N-methylmorpholine were added. Thereaction mixture was stirred for 3 hours at 0° C. under nitrogen andmost of the DMF was subsequently distilled off at a bath temperature of35° C. and −1 mbar. The residue was purified by column chromatography(eluant: CH₂Cl₂/MeOH, 45/5, toward the end with addition of 0.7 parts of50% strength acetic acid). This gave 0.75 g of a slightly yellowishamorphous powder.

The latter was dissolved in 5 ml of CH₂C1₂ and 10 ml of trifluoroaceticacid and the solution was left to stand overnight at room temperature.After addition of 30 ml of toluene, the mixture was concentrated invacuo, and the residue was treated with ether and subsequently convertedinto the betain on a silica gel column (eluant: MeOH/25% strength NR₃,50/2). The betain was dissolved in 20 ml of water, brought to pH 4.5with 1N HCl and lyophilized. This gave 0.36 g of amorphous powder,FAB-MAS (M+H⁺): 476.

EXAMPLE 65N-(Hydroxycarbonylmethylene)-(R)-cyclohexylalanyl-(3S)-pyrazolidine-3-carboxylicAcid 2-(4-Amidino-thienylmethyl)amide Hydrochloride

8.36 g (21.7 mmol) ofN-Boc-N-(tert-butoxycarbonylmethylene)-(D)-cyclohexylalanine and 5 g(21.7 mmol) of methyl (3S)-1-tert-butoxycarbonylpyrazoline-3-carboxylate[H. O. Kim, C. Lum, M. S. Lee (1997), THL 38 (28), 4935] were dissolvedin 60 ml of CH₂Cl₂, 6.1 g (31.8 mmol) of EDC.HCl were added withstirring at −80C, and, after a further 20 minutes, 4.0 g (31 mmol) ofdiisopropylethylamine were added. After the mixture had been stirred for40 minutes, 0.8 g of DMAP was added and the mixture was left to standfor 2 days at room temperature. After addition of 200 ml of ether, themixture was washed with 5% strength citric acid, 5% strength NaHCO₃solution and water, and, after drying, the ether was distilled off.After purification by column chromatography (eluant: CH₂Cl₂/acetone,50/2), 8.2 g (63% of theory) of a white, amorphous powder were isolated.

Hydrolysis: 8.0 g (13.4 mmol) of the ester were dissolved in 60 ml ofdioxane and 12 ml of water, and 15 ml of 1N NaOH were added at 10° C.After 1.5 hours, the pH was brought to 8 with 1N HCl, the dioxane wasdistilled off, and the residue was diluted with 250 ml of water andextracted with ether. The aqueous phase was brought to pH 2.5 with 1NKHSO₄ solution, and the acid which had separated out was extracted withether. After the ether had been stripped off, 7.7 g of amorphous acidremained. A sample recrystallized from water-saturated n-hexane melts at115 to 120° C. and has an angle of rotation [α]_(D) ²⁰ of +112.4° C.(CHCl₃, c=1).

The coupling with 4-aminomethyl-2-amidinothiophene dihydrochloride wascarried out by a method similar to that described in Example 64 step b).After purification by column chromatography (eluant: CH₂Cl₂/MeOH/50%strength acetic acid, 40/10/0.7), 4 g ofN-Boc-N-(tert-butoxycarbonylmethylene)-(R)-cyclohexylalanyl-(3S)-pyrazolidine-3-carboxylicacid [4-(2-amidino)thienylmethyl]amide acetate were obtained startingfrom 4.15 g of the above acid.

Cleavage [sic] of the protective group: The above compound was dissolvedin 12 ml of dioxane, 20 ml of 1N HCl were added, and the mixture washeated for 4.5 hours at 75° C. The solution was diluted with 50 ml ofwater, brought to pH 4 with an ion exchanger (3-A4 resin, BioRad), andthe water was distilled off. The residue was dissolved in isopropanoland the hydrochloride was precipitated by addition of ether. Afterpurification by column chromatography (eluant: CH₂Cl₂/MeOH/50% strengthacetic acid, 35/15/7), the residue was dissolved in water, brought to pH4 with 1N HCl and lyophilized. This gave 1.6 g of the amorphoushydrochloride, FAB-MS (M+H⁺): 465.

The following were obtained by a method similar to that described inExample 65:

EXAMPLE 66N-(Hydroxycarbonylmethylene)-(R)-cyclohexylalanyl-(3R)-pyrazolidine-3-carboxylicAcid [4-(2-Amidino)thienylmethyl]amide Hydrochloride White AmorphousPowder. FAB-MS (M+H⁺): 465 EXAMPLE 67N-(Hydroxycarbonylmethylene)-(R)-cyclohexylglycyl-(3R)-pyrazolidine-3-carboxylicAcid [4-(2-Amidino)-thienylmethyl]amide Hydrochloride White AmorphousPowder. FAB-MS (M+H⁺): 451 EXAMPLE 68N-(Hydroxycarbonylmethylene)-(R)-cyclohexylglycyl-(3S)-pyrazolidine-3-carboxylicAcid [4-(2-Amidino)-thienylmethyl]amide Hydrochloride White AmorphousPowder. FAB-MS (M+H⁺): 451 EXAMPLE 69N-(Hydroxycarbonylmethylene)-((R)-cyclohexylglycyl-(−)-thiazolidine-2-carboxylicAcid [4-(2-Amidino)thienylmethyl]amide Hydrochloride Starting Material:Methyl (−)-Thiazolidine-2-carboxylate [R. L. Johnson, E. E. Smissman(1978), J. Med. Chem. 21, 165] EXAMPLE 70N-(Hydroxycarbonylmethylene)-(R)-cyclohexylalanyl-(−)-thiazolidine-2-carboxylicAcid [4-(2-Amidino)thienylmethyl]amide White Amorphous Powder. FAB-MS(M+H⁺): 482 EXAMPLE 71N-(Hydroxycarbonylmethylene)-(R)-cyclohexylalanyl-(L)-octahydroindole-2-carboxylicAcid [4-(2-Amidino)-thienylmethyl]amide Hydrochloride White AmorphousPowder. FAB-MS (M+H⁺): 518 EXAMPLE 72N-(Hydroxycarbonylmethylene)-(R)-cyclohexylglycyl-(L)-octahydroindole-2-carboxylicAcid [4-(2-Amidino)-thienylmethyl]amide Hydrochloride White AmorphousPowder. FAB-MS (M+H⁺): 504 EXAMPLE 73N-(Hydroxycarbonylmethylene)-(D)-cyclohexylalanyl-(45)-5.5-dimethylthiazolidine-4-carboxylic[sic] Acid [2-(4-Amidino)thienylmethyl]amide

This compound can be prepared by the following reaction sequence:coupling of N-(t-BuO₂C—CH₂)-N-Boc-(D)-Cha-OH with (5)-5.5-Me₂-thz-4-OMe-[sic] [J. Samanen u.a. (1990), Int. J. Peptide Protein Res. 35, 501(1990)] to give N-(t-BuO₂C—CH₂)-Boc-(D)-Cha-(2,2-Me₂-thz-4)-OMe,alkaline hydrolysis of the methyl ester, coupling of the resulting acidwith H₂N—CH₂-(2-CN)-2-thioph to giveN-(t-BuO₂C—CH₂)-N-Boc-(D)-Cha-(2.2-Me₂-thz-4)-NE-CH₂-2-(4-CN)-thioph[sic], amidine formation and subsequent elimination of the protectivegroups by a method similar to that described in Example 2b-e. FAB-MS(M+H⁺): 505; m.p. 184-7° C. (decomp.)

EXAMPLE 74N-(Hydroxycarbonylmethylene)-(D)-cyclohexylglycyl-5,5-dimethylthiazolidine-4-carboxylicAcid [4-(2-Amidino)thienylmethyl]amide

This compound can be prepared by the following reaction sequence:coupling of N-(t-BuO₂C—CH₂)-N-Boc-(D)-Chg-OH 10 with 5.5-Me₂-thz-4-OMe-[sic] to give N-(t-BuO₂C—CH₂)-N-Boc-(D)-Chg-(2,2-Me₂-thz-4)-OMe,alkaline hydrolysis of the methyl ester, coupling of the resulting acidwith H₂N—CH₂-(2-CN)-4-thioph to giveN-(t-BuO₂C—CH₂)-N-Boc-(D)-Chg-(5.5-Me₂-thz-4)-NH—CH₂-4-(2-CN)-thioph[sic], amidine formation and subsequent elimination of the protectivegroups by a method similar to that described in Example 2b-e. FAB-MS(M+H⁺): 491, m.p. 164-166° C. (decomp.)

EXAMPLES 75-90

The following compounds can be synthesized from the corresponding A—B—D—and E—F— units using a method similar to that described in Example 14 inWO98/06741:

75. HOOC—CH₂-(D)-(p-OMe-Phe)-Pro-NH—CH₂-2-(4-am)-thioph

76. HOOC—CH₂-(D)-(p-OMe-m-Cl-Phe)-Pro-NH—CH₂-2-(4-am)-thioph

77. HOOC—CH₂-(D)-(p-OMe-Phe)-Pro-NH—CH₂-5-(2-am-3-Me)-thioph

78. HOOC—CH₂-(D)-(p-CF₃-Phe)-Pro-NH—CH₂-5-(2-am-3-Me)-thioph

79. HOOC—CH₂-(D)-(p-Cl-m-Cl-Phe)-Pro-NH—CH₂-5-(2-am-3-Me)-thioph

80. HOOC—CH₂-(D)-(p-CF₃-Phe)-Pro-NH—CH₂-2-(4-am)-thioph

81. HOOC—CH₂-(D)-Cha-Pro-NH—CH₂-2-(4-am-5-Me)-thiaz

82. HOOC—CH₂-(D)-Chg-Pro-NH—CH₂-2-(4-am-5-Me)-thiaz

83. HOOC—CH₂-(D)-(p-OMe-Phe)-Pro-NH—CH₂-2-(4-am-5-Me)-thiaz

84. HOOC—CH₂-(D)-(p-OMe-Phe)-Pro-NH—CH₂-2-(4-am)-thiaz

85. HOOC—CH₂-(D)-(p-CF₃-Phe)-Pro-NH—CH₂-2-(4-am)-thiaz

86. HOOC—CH₂-(D)-(p-CF₃-Phe)-Pro-NH—CH₂-2-(4-am-5-Me)-thiaz

87. HOOC—CH₂-(D)-(p-iPr-m-Me-Phe)-Pro-NH—CH₂-2-(4-am)-thiaz

88. HOOC—CH₂-(D)-(p-OMe-m-Cl-Phe)-Pro-NH—CH₂-2-(4-am)-thiaz

89. HOOC—CH₂-(D)-(p-Cl-m-Cl-Phe)-Pro-NH—CH₂-2-(4-am)-thiaz

90. HOOC—CH₂-(D)-(p-Cl-m-Cl-Phe)-Pro-NH—CH₂-2-(5-am-4-Me)-thiaz

EXAMPLE 91

The following compound can be synthesized from the corresponding E— unitand N-(t-BuO₂C—CH₂)-N-Boc-(D)-Cha-Pro-OH by a method similar to thatdescribed in Example 20:

HOOC—CH₂-(D)-Cha-Pro-NH—CH₂-3-(5 am-1-Me)-pyraz

EXAMPLE 92

HOOC—CH₂-(D)-Cha-Thz-4-NH—CH₂-5-(2-am)-thioph was synthesized fromBoc-Thz-4-OH, 5-H₂N—CH₂-thioph-2-CN andN-(t-BUO₂C—CH₂)-N-Boc-(D)-Cha-Pro-OH by a method similar to thatdescribed in Example 3.

PHARMACOLOGICAL EXAMPLES EXAMPLE A

Chromogenic Test for Kallikrein Inhibitors

Reagents: Human plasma kallikrein (No. K 3126, Sigma, Deisenhofen,Germany)

Substrate: Chromozym GK (No. 709875, Boehringer, Mannheim, Germany)

Buffer: 20 mM Tris(HCl [sic] pH=8.50

Experimental Procedure:

Chromogenic test for determining the kallikrein activity is carried outin microplates. 2 gl of the solution of the substance in DMSO are addedto 93 μl of buffer, and this is mixed with a final concentration of 0.01units/ml kallikrein. Incubation is for 10 minutes at 20 to 25° C. Thetest is started by adding 100 μl of substrate (500 μmol/l finalconcentration). After incubation for a further 30 minutes, theabsorption is measured in a photometer at 405 nm.

EXAMPLE B Thrombin Time

Reagents: Thrombin Reagent (Product No. 126 594, Boehringer, Mannheim,Germany)

Preparation of the Citrate Plasma:

9 parts of venous human blood from the vena cephalica are mixed with onepart of sodium citrate solution (0.11 mol/l). The mixture issubsequently centrifuged. The plasma can be stored at −20° C.

Experimental Procedure:

50 μl of the solution of the test substance and 50 μl of citrate plasmaare incubated for 2 minutes at 37° C. (CL8, ball type, Bender & Hobein,Munich, FRG). 100 μl of thrombin reagent (37° C.) are subsequentlyadded. The time that elapses until the fibrin clot forms is determined.

EXAMPLE C Chromogenic Test for Thrombin Inhibitors

Reagents: Human Plasma Thrombin (No. T-8885, Sigma, Deisenhofen,Germany)

Substrate: H-D-Phe-Pip-Arg-pNA2HCl (S-2238, Chromogenix, M6lndahl,Sweden)

Buffer: Tris 50 mmol/l, NaCl 154 mmol/l, pH 8.0

Experimental Procedure:

The chromogenic test can be carried out in microtiter plates. 10 μl of asolution of the substance in DMSO are added to 250 μl of buffer withthrombin (final concentration 0.1 NIH units/ml) and the mixture isincubated for 5 minutes at 20 to 28° C. The test is started by adding 50μl of the solution of the substrate in buffer (final concentration 100μmol/l), and the mixture is incubated at 28° C. and, after 5 minutes,the reaction is stopped by adding 50 μl of citric acid (35% strength).The absorption is measured at 405/630 nm.

EXAMPLE D Platelet Aggregation in Platelet-rich Plasma

Reagents: Human Plasma Thrombin (No. T-8885, Sigma, Deisenhofen,Germany)

Preparation of the Citrate-rich Platelet-rich Plasma:

Venous blood is collected from vena cephalica from healthy unmedicatedsubjects. The blood is mixed 9:1 with 0.13-molar trisodium citrate.

Platelet-rich plasma (PRP) is prepared by centrifugation at 250×g (10minutes at room temperature). Platelet-poor plasma (PPP) is prepared bycentrifugation for 20 minutes at 3600×g. PRP and PPP can be stored for 3hours at room temperature in closed PE vessels. The plateletconcentration is measured with a hematocytometer and should be between2.5 and 2.8.10⁸/ml.

Experimental Procedure:

The platelet aggregation is measured turbidimetrically at 37° C. (PAP 4,Biodata Corporation, Horsham, PA, USA). Before thrombin is added, 215.6μl of PRP are incubated for 3 minutes with 2.2 μl of test substance andthe mixture is then stirred for 2 minutes at 1000 rpm. At a finalconcentration of 0.15 NIH units/ml, 2.2 μl of thrombin solution lead tothe maximum aggregation effect at 37° C./1000 rpm. The inhibitory effectof the test substances is determined by comparing the rate at whichthrombin aggregates (slope) without substance with the rate of thrombinwith test substance at various concentrations.

We claim:
 1. A compound of the formula I A—B—D—E—F  I in which A, B, D,E and F have the following meanings: A:

 where m is 0, 1 or 2, n is0, 1 or 2, R¹ is HOOC—, C₁₋₆-alkyl-OOC—,aryl-C₀₋₄-alkyl-OOC or —OH, R² is H—, C₁₋₄-alkyl- or R¹—(CH₂)_(m)— andR³ is H— or C₁₋₄-alkyl-, B:

 where R⁴ is H—, C₁₋₄-alkyl- or R¹—(CH₂)_(m)—(where R¹ and m have theabovementioned meanings), p is 0 or 1, R⁵ is H— or C₁₋₄-alkyl-, R⁶ isH—, C₁₋₈-alkyl-, 2-thienyl-, 3-thienyl-, 3-indolyl-, 4-imidazolyl-,2-pyridyl-, 3-pyridyl-, 4-pyridyl-, phenyl- which may carry up to threeidentical or different radicals selected from the group of C₁₋₄-alkyl-,CF₃—, C₁₋₄-alkoxy-, HO—, BnO—, F— and Cl—, C₃₋₈-cycloalkyl- which maycarry up to four identical or different C₁₋₄-alkyl- radicals and/orwhere one or two C—C single bonds in the ring can be replaced by a C═Cdouble bond and/or a phenyl ring can be fused on, C₇-C₁₂-bicycloalkyl-or C₁₀-tricycloalkyl- or R⁴ and R⁶ together are an ethylene or propylenegroup, R⁷ is H, C₁₋₈-alkyl-, phenyl- which may carry up to threeidentical or different radicals from the group of C₁₋₄-alkyl-, CF₃—,C₁₋₄-alkoxy-, F— and Cl—, or C₃₋₈-cycloalkyl- which may carry up to fouridentical or different C,₄-alkyl radicals, and R⁸ is H or C₁₋₄-alkyl, D:

where R²⁰ is H, C₁₋₄-alkyl, Bnor BnO(CO)— and where the followingapplies: if D is II, III or XI, then E has the following meaning:

where a) in the event that X=S, O, NH or NR¹², Y is —CR¹³═, —CH═ and Zis —CR═ or Y is —CR³═ and Z is —CH═ or b) in the event that X=NR¹², Y is—CH═ and Z is —CH═ or c) in the event that X=S, O or NH, Y is —CR¹⁵═ andZ is —N═ or Y is —N═ and Z is —CR¹⁵ ═ or d) in the event that X=—NR¹²—,Y is —N═ and Z is —CR¹⁶═, —N═ or Y is —CR¹⁶═ and Z is —N═ and R⁹ is H—or C₁₋₃-alkyl-, R¹⁰ is H— or C₁₋₄-alkyl-, R¹¹ is H— or C₁₋₄-alkyl-, R¹²is CH₃— or C₂H₅—, R¹³ is Cl—, CF₃— or C₁₋₄-alkyl-, R¹⁴ is Cl—, CF₃— orC₁₋₄-alkyl-, R¹⁵ is CF₃— or C₁₋₄-alkyl-, R¹⁶ is H—, CF₃— or C₁₋₄-alkyl-and R²⁰ is as above, or, if D is IV, VI, VII, VIII, IX or X, then E hasthe following meaning:

where X is O, S or —NR¹⁷— and Y is —N═ and Z is —CR¹⁶═ or —N═ or Y is—CR¹⁶═ and Z is —N═ or Y is —CR¹⁸═ and Z is —CR¹⁹═ and R⁹, R¹⁰, R¹¹, R¹⁶and R²⁰ are as above, R¹⁷ is H, CH₃— or C₂H₅—, R¹⁸ is H—, Cl—, CF₃— orC₁₋₄-alkyl-, R¹⁹ is H—, Cl—, CF₃— or C₁₋₄-alkyl-, or if D is II, III,IV, VI, VII, VIII, IX, X or XI, E has the following meanings:

where a) in the event that X=S, Y is —CR¹⁸═ and Z is  CR¹⁹═ or Y is—CR¹⁶═ and Z is —N═ or b) in the event that X=O or —NR¹²—, Y is —N═,—CR¹⁶═ and Z is —N═, —CR¹⁸═ and R⁹, R¹⁰, R¹¹, R¹², R¹⁶, R¹⁸, R¹⁹ and R²⁰have the abovementioned meanings, F:

or a salt thereof with a physiologically acceptable acid.
 2. A compoundof the formula I as claimed in claim 1, where A to E have the followingmeanings: A: HOOC—(CH₂)_(t)—(t=1, 2 or 3), (HOOC—CH₂)₂—CH—,HOOC—CH₂—CH(COOH)—, HOOC—CH(C₁₋₄-alkyl)-, HOOC—C(C₁₋₄-alkyl)₂—,C₁₋₆-alkyl-OOC—(CH₂)_(t)—,

p is 0 or 1, R⁴ is H—, C₁₋₄-alkyl- or HOOC—(CH₂)_(m)— (m=1, 2 or 3), R⁵is H—, methyl- R⁶ is H—, C₁₋₈-alkyl-, 2-thienyl-, 3-thienyl-,3-indolyl-, 4-imidazolyl-, 2-pyridyl-, 3-pyridyl-, 4-pyridyl-, phenyl-which may carry up to three identical or different radicals from thegroup of CH₃—, CF₃—, CH₃—O—, HO—, BnO—, F— and Cl—, C₃₋₈-cycloalkyl,which may carry up to four methyl radicals, bicyclooctyl-,bicycloheptyl-, adamantly-, indanyl-, or decalinyl-, R⁷ is H,C₁₋₈-alkyl-, phenyl-, which may carry up to three identical or differentradicals from the group of CH₃—, CF₃—, CH₃O—, F— or Cl—, orC₃₋₈-cycloalkyl- which may carry up to four methyl radicals, R⁸ is H,C₁₋₄-alkyl, D:

where R²⁰ is H, CH₃, Bn or BnO(CO)— and where the following applies: ifD is II, III or XI, then E has the meaning:

where a) in the event that X=S, O, NH or NR¹⁷, Y is —CR¹³═ or —CH═ and Zis —CR¹⁴═ or Y is —CR¹³═ and Z is —CH═ or b) in the event that X=NR¹², Yis —CH═ and Z is —CH═ or c) in the event that X=S, O or NH, Y is —CR¹⁵═and Z is —N═ or Y is —N═ and Z is —CR¹⁵═ or d) in the event that X=NR¹²,Y is —N═ and Z is —CR¹⁶— or —N═ or Y is —CR¹⁶═ and Z is —N═ and R¹² isCH₃— or C₂H₅—, R¹³ is Cl—, CF₃— or C₁₋₄-alkyl-, R¹⁴ is Cl—, CF₃— orC₁₋₄-alkyl-, R¹⁵ is CF₃— or C₁₋₄-alkyl-, R¹⁶ is H—, CF₃— or C₁₋₄-alkyl-and R¹⁷ is H, CH₃— or C₂H₅—, R²⁰ is as above, or if D is IV, VI, VII,VIII, IX or X, then E has the meaning:

where X is O, S or —NR¹⁷— and where Y is —N═ and Z is —CR¹⁶═ or —N═ or Yis —CR¹⁶═ and Z is —N═ or Y is —CR¹⁸═ and z is —CR¹⁹═ and R¹⁶, R¹⁷, andR²⁰ have the abovementioned meanings, R ¹⁸ is H—, Cl—, CF₃— orC₁₋₄-alkyl-, and R¹⁹ is H—, Cl—, CF₃— or C₁₋₄-alkyl-, or if D is II,III, IV, VI, VII, VIII, IX, X or XI, then E has the meanings:

where a) in the event that X=S, Y is —CR¹⁸═ and Z is —CR¹⁹═ or Y is—CR¹⁶═ and Z is —N═ or b) in the event that X=O or —NR¹²—, Y is —N═ or—CR¹⁶═ and Z is —N═ or —CR¹⁸═ and R¹², R¹¹, R¹⁸, R¹⁹ and R²⁰ have theabovementioned meanings, F:

and their salts with physiologically acceptable acids.
 3. A compound ofthe formula I as claimed in claim 1, where A, B, D, E and F have thefollowing meanings: A: HOOC—CH₂, HOOC—CH₂—CH₂, HOOC—CH(CH₃),HOOC—CH(C₂H₅) B:

p is 0 or 1, R⁴ is H—, CH₃— R⁵ is H—, CH₃—, R⁶ is C₁₋₈-alkyl-,C₅₋₈-cycloalkyl- which may carry up to four methyl radicals, 2-thienyl-,3-indolyl-, 4-imidazolyl-, 2-pyridyl-, 3-pyridyl-, 4-pyridyl, phenyl-which may carry up to three identical or different radicals from thegroup of CH₃—, CF₃—, CH₃O—, HO—, BnO—, F— and Cl—, bicyclooctyl,bicycloheptyl, adamantly, indanyl, or decalinyl, R⁷ is H, CH₃—, R⁸ is H,CH₃—, D:

where R²⁰ is H, BnO(CO)— and where the following applies: if D is II,III or XI, then E has the meaning:

where X is —S— and where Y is —CH═ and Z is —CR³═ or Y is —CR¹³═ and Zis —CH═ or Y is —CR¹⁵═ and Z is —N═ or Y is —N═ and Z is —CR¹⁵═ and R¹³is Cl—, CF₃— or CH₃— R¹⁵ is CF₃— or CH₃— and R²⁰ is as above, or if D isIV, VI, VII, VIII, IX or X, then E has the meaning:

where X is S and where Y is —N═ and Z is —CR¹⁶═ or Y is —CR¹⁶═ and Z is—N═ or Y is —CR¹³═ and Z is —CH═ or Y is —CH═ and Z is —CR¹³═ or Y is—CH═ and Z is —CH═ and R¹³, R²⁰ have the abovementioned meanings, R¹⁶ isH—, CF₃— or CH₃— or if D is II, III, IV, VI, VII, VIII, IX, X or XI,then E has the meanings:

where either a) in the event that X=S, Y is —CH═ and Z is —CR¹⁸═ or Y is—CR¹⁶═ and Z is —N═ or Y is —CR¹⁸═ and Z is —CH═ or b) in the event thatX=O or —NCH₃ Y is —CH═ and Z is —CR¹⁶═ or Y is —CR¹⁶═ and Z is —CH═ orc) in the event that X=—NR¹²— Y is —N═ and Z is —CR¹⁸═ and R¹² is CH₃—or C₂H₅— and R¹⁸ is H, Cl—, CF₃— or CH₃—, and R¹⁶, R²⁰ have theabovementioned meanings F:

and their salts with physiologically acceptable acids.
 4. A compound ofthe formula I as claimed in claim 1, where A, B, D, E and F have thefollowing meanings: A: HOOC—CH₂, HOOC—CH₂—CH₂, HOOC—CH(CH₃),HOOC—CH(C₂H₅) B:

p is 0 or 1, R⁴ is H—, R⁵ is H—, R⁶ is C₁₋₈-alkyl-, 2-thienyl-,3-indolyl-, 4-imidazolyl-, 2-pyridyl-, 3-pyridyl-, 4-pyridyl-,C₅₋₈-cycloalkyl- which may carry up to four methyl radicals, phenyl-which may carry up to three identical or different radicals from thegroup of CH₃—, CF₃—, CH₃O—, HO—, BnO—, F—and Cl—, bicyclooctyl,bicycloheptyl, adamantly, indanyl, or decalinyl, R⁷ is H, R⁸ is H, D:

where the following applies: if D is II, III or XI, then E has themeaning:

where X is S and Y is —CR¹³═ and Z is —CH═ or is —CH═ and Z is —CR¹³═ orY is —CR¹⁵═ and Z is —N═ or Y is —N═ and Z is —CR¹⁵═ and R¹³ is Cl—,CF₃— or CH₃— and R¹⁵ is CF₃— or CH₃—, or if D is IV, VI, VII, VIII, IXor X, then E has the meaning:

where X is S and Y is —N═ and Z is —CR¹⁶═ or Y is —CR¹⁶═ and Z is —N═ orY is —CH═ and Z is —CR¹³═ or Y is —CR¹ ³═ and Z is —CH═ or Y is —CH═ andZ is —CH═ and R¹³ has the abovementioned meaning and R¹⁶ is H—, CF₃— orCH₃—, or if D is II, III, IV, VI, VII, VIII, IX, X or XI, then E has themeanings:

where a) in the event that X=S, Y is —CH═ and Z is —CR¹⁸═ or Y is —CR¹⁸═and Z is —CH═ or Y is —CR¹⁶═ and Z is —N═ or b) in the event that X=O or—NCH₃ Y is —CH═ and Z is —CR¹⁶═ or Y is —CR¹⁶═ and Z is —CH═ or c) inthe event that X=NCH₃ Y is —N═ and Z is —CR¹⁶═ and R¹⁶ has theabovementioned meaning and R¹⁸ is H, Cl—, CF₃— or CH₃—, F:

and their salts with physiologically acceptable acids.
 5. A compoundwhich contains the structural element

where D and E have the meanings given in any of claims 1 to 4 and wherea hydrogen atom, a protective group, an unsubstituted or substitutednatural or unnatural amino acid, an unsubstituted or substitutedcarboxylic acid or an unsubstituted or substituted alkyl radical islocated on the nitrogen atom of building block D.
 6. A compound whichcontains the structural element

where E has the meaning given in any of claims 1 to 4 and where ahydrogen atom, a protective group, an unsubstituted or substitutednatural or unnatural amino acid, an unsubstituted or substitutedcarboxylic acid or an unsubstituted or substituted alkyl radical islocated on the nitrogen atom of NR⁹.
 7. A compound containing astructural element of the formula

where Q is CH₃ or Cl, T is NCH₃, O or S and W is NCH₃ or S.
 8. Acompound of the formula Va or Vb A—B—D—E—CN  Va, A—B—D—E—CSNH₂  Vb,where A, B, D and E have the meanings given in claim
 1. 9. A drugcomprising, in addition to drug adjuvants and/or excipients, a compoundas defined in claim 1 and a compound which contains the structuralelement

where D and E have the meanings given in claim
 1. 10. A process forcombating a disease selected from the group consisting of deep veinthrombones, pulmonary embolisms, myocardial or cerebral infarcts,unstable angina, disseminated intravascular coagulation, lengthenedrenerfusion time and shortened reocclusion time in a patient in needthereof which comprises treating the patient with an effective amount ofa drug as defined in claim
 1. 11. A method for reducing coagulation inextracorporeal circulation, comprising coating an artificial surface ora tubing system or line of a machine used for this purpose with acompound of the formula I as claimed in claim
 1. 12. A process forinhibiting thrombin in a patient in need of such treatment, comprisingintroducing an effective amount of a compound as defined in claim 1 suchthat the compound interacts with and inhibits the activity of thrombin.13. A process for combating an inflammatory disease selected from thegroup consisting of asthma, pancreatitis, rhinitis, arthritis andurticaria in a pateient in need thereof which comprises treating thepatient with an effective amount of a compound of claim 1 and a compoundwhich contains the structural element

where D and E have the meanings given in claim 1 and where a hydrogenatom, a protective group, an unsubstituted or substituted amino acid, anunsubstituted or substituted carboxylic acid or an unsubstituted orsubstituted alkyl radical is located on the nitrogen atom of buildingblock D.
 14. A process for producing drugs which are suitable asthrombin inhibitors, comprising combining a compound as claimed in claim1 with drug adjuvants and/or excipients.